1,3,5-Tris[(1E)-2′-(4′′-benzoic acid)vinyl]benzene] (Ramizol™)

TSB-007

1,3,5-Tris[(1E)-2′-(4′′-benzoic acid)vinyl]benzene] (Ramizol™) is a potent and non-toxic synthetic antimicrobial agent, and we now establish that it is also a potent inhibitor of reactive oxygen species (ROS) generation, with similar antioxidant activity to α-tocopherol (Vitamin E), which is a standard antioxidantdrug.

Ramizol, useful for treating bacterial infections such as Gram positive bacterial infection. Boulos & Cooper Pharmaceuticals could be seen to have ramizol in preclinical development for treating Clostridium difficile associated diseases.  preparation of ramizol that was first described by the inventor Dr Ramiz Boulos, one of the company’s founding directors and CEO, in WO2011075766 as TSB-007 (claim 3, page 71) – said family of patenting having been originally assigned to the University of Western Australia and from whom Dr Boulos is reported to have acquired the rights to said intellectual property in late 2012 (ramizol having seemingly been previously being developed by the University with the name NAL-135B for treating Gram positive bacterial infections).

Professor Ramiz Boulos with a vial of Ramizol

A scientific paper released today in the Journal of Antibiotics presents the pre-clinical development of Ramizol®, a first generation drug belonging to a new class of styrylbenzene antibiotics with a novel mechanism of action.

The research was undertaken by Australian company Boulos & Cooper Pharmaceuticals in partnership with the University of South Australia, Flinders University, Eurofins Panlabs and Micromyx LLC. The study found that over 99.9% of the drug, administered orally, stays in the gastrointestinal tract where it can reach the bacteria in the colon at high enough concentrations to yield a therapeutic effect.

Chief Executive Officer of Boulos & Cooper Pharmaceuticals, Dr Ramiz Boulos, said “this new class of antibiotics has antioxidant properties and can be manufactured for a low cost; benefits that will be felt by the end-user”.

The new antibiotic has low frequency of resistance and shows promise as a monotherapy for the treatment of Clostridium difficile associated disease. Dr Boulos stated “we are very excited about these results given the unforgiving nature of Clostridium difficile infections”. He added “In a world where there are few treatment options, we are desperate for new antibiotics to fight intractable infections”.

The company expects to start Phase I clinical trials in 2017.

1,3,5-Tris[(1E)-20 -(400-benzoic acid)vinyl]benzene……………….recrystallised from THF/H2O and dried to give the triacid as a pale brown powder.

1 H NMR (500.1 MHz, d6-DMSO): d 7.49 (m, 6H, vinyl CH), 7.76 (d, J 8.5, 6H, ArH), 7.88 (s, 3H, core ArH), 7.98 (d, J 8.5, 6H, ArH);

13C NMR (125.8 MHz, d6-DMSO): d [ppm] 125.0, 126.5, 128.4, 129.7, 129.9, 130.50, 137.6, 141.3, 167.1;

IR (KBr): n [cm1 ] 3067, 3026, 1684 (nC¼O), 1604, 1566, 1420, 1384, 1312, 1286, 1179;

HR-EIþ-MS: C33H24O6 requires 516.1573 amu, found 516.1564;

EIþ-MS: MI ¼ C33H24O6; m/z: 516.1 (100%) ¼ MIþ, 472.1 (11.3%) ¼ [MI CO2] þ.

The Synthesis of Fluorescent DNA Intercalator Precursors through Efficient Multiple Heck Reactions

Nigel A. Lengkeek ^A , Ramiz A. Boulos ^A , Allan J. McKinley ^A , Thomas V. Riley ^C , Boris Martinac ^B and Scott G. Stewart ^{A D}

PATENT

 WO 2011075766

PATENT

WO-2017027933

Compounds with antimicrobial properties have attracted great interest in recent times as a result of an increase in the prevalence of infections caused by Gram-positive bacteria, resulting in serious or fatal diseases. Furthermore, the regular use of broad spectrum antibiotic formulas has led to the increased occurrence of bacterial strains resistant to some antimicrobial formulations.

Novel antimicrobial compounds have the potential to be highly effective against these types of treatment-resistant bacteria. The pathogens, having not previously been exposed to the antimicrobial formulation, may have little to no resistance to the treatment.

International patent application WO 2012/075766 describes a series of novel aryl compounds and their use as antimicrobials to treat bacterial infections or diseases. The chemical synthesis of a therapeutic drug has a direct effect on its cost, dosing regimens and popularity. Drugs with complicated or expensive chemical synthesis will find it challenging to reach the market, notwithstanding their efficacy. Further, syntheses amenable to application at commercial scales are highly advantageous. The development of an efficient and large-scale synthesis of a therapeutic drug is critical for its drug developmental pathway, and highly commercially advantageous.

1H NMR PREDICT

13C NMR PREDICT

REFERENCES

N. A. Lengkeek, R. A. Boulos, A. J. McKinley, T. V. Riley, B. Martinac and S. G. Stewart, Aust. J. Chem., 2011, 64, 316–323

http://pubs.rsc.org/en/content/articlehtml/2013/ra/c3ra40658j#cit11

/////////////Ramizol, PHASE 1, TSB-007

OC(=O)c4ccc(/C=C/c3cc(/C=C/c1ccc(cc1)C(=O)O)cc(/C=C/c2ccc(cc2)C(=O)O)c3)cc4

ASP ?

(2R)-2-(4-cyclopropanesulfonyl-3-cyclopropylphenyl)-N-[5-(hydroxymethyl)pyrazin-2-yl]-3-[(R)-3-oxocyclopentyl]propanamide

Synthesis

contd…………………………..

PATENT

WO2009091014

https://patentscope.wipo.int/search/en/detail.jsf;jsessionid=56E9927692EF5105140FE1CD1FD14A5D.wapp1nC?docId=WO2009091014&recNum=114&maxRec=374&office=&prevFilter=&sortOption=&queryString=FP%3A%28astellas+pharma%29&tab=FullText

A Practical and Scalable Synthesis of a Glucokinase Activator via Diastereomeric Resolution and Palladium-Catalyzed C–N Coupling Reaction

Yohei Yamashita^*†§ , Yasuhiro Morinaga^†, Makoto Kasai^†, Takao Hashimoto^†, Yuji Takahama^†, Atsushi Ohigashi^†, Satoshi Yonishi^‡, and Motohiro Akazome^§

^† Process Chemistry Labs., Astellas Pharma Inc., 160-2 Akahama, Takahagi-shi, Ibaraki 318-0001, Japan

^‡Astellas Research Technologies Co., Ltd., 21 Miyukigaoka, Tsukuba-shi, Ibaraki 305-8585, Japan

^§ Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, Chiba University, 1-33 Yayoicho, Inageku, Chiba 263-8522, Japan

Org. Process Res. Dev., Article ASAP

DOI: 10.1021/acs.oprd.6b00415

 

Here we describe the research and development of a process for the practical synthesis of glucokinase activator (R)-1 as a potential drug for treating type-2 diabetes. The key intermediate, chiral α-arylpropionic acid (R)-2, was synthesized in high diastereomeric excess through the diasteromeric resolution of 7 without the need for a chiral resolving agent. The counterpart 2-aminopyrazine derivative 3 was synthesized using a palladium-catalyzed C–N coupling reaction. This efficient process was demonstrated at the pilot scale and yielded 19.0 kg of (R)-1. Moreover, an epimerization process to obtain (R)-7 from the undesired (S)-7 was developed.

Hayakawa, M.; Kido, Y.; Nigawara, T.; Okumura, M.; Kanai, A.; Maki, K.; Amino, N. PCT Int. Appl. WO/2009/091014 A1 20090723,2009.

https://www.astellas.com/en/ir/library/pdf/3q2017_rd_en.pdf

///////////1174229-89-0, ASTELLAS, Glucokinase Activator, TYPE 2 DIABETES, PRECLINICAL, ASP ?, WO 2009091014, Masahiko Hayakawa, Yoshiyuki Kido, Takahiro Nigawara, Mitsuaki Okumura, Akira Kanai, Keisuke Maki, Nobuaki Amino, WO2009091014,

O=C(Nc1cnc(cn1)CO)[C@H](C[C@@H]2CC(=O)CC2)c3ccc(c(c3)C4CC4)S(=O)(=O)C5CC5

Troxacitabine

CAS 145918-75-8

• Molecular FormulaC₈H₁₁N₃O₄
• Average mass213.191 Da

Hmd-cytosine; NCGC00183848-01; Beta-L-Dioxolane-cytidine; 4-amino-1-[(2S)-2-(hydroxymethyl)-1,3-dioxolan-4-yl]pyrimidin-2-one; 2R(-)-cis-Hmd-cytosine, (-)-ODDC

4-amino-1-[(2S)-2-(hydroxymethyl)-1,3-dioxolan-4-yl]pyrimidin-2-one

Troxacitabine (brand name Troxatyl) is a nucleoside analogue with anticancer activity. Its use is being studied in patients with refractory lymphoproliferative diseases.^[1]

Troxacitabine (brand name Troxatyl) is a nucleoside analogue with anticancer activity. Its use is being studied in patients with refractory lymphoproliferative diseases.

PATENT

https://www.google.com/patents/WO1992018517A1?cl=en

WO 9218517

SYNTHESIS

WO 2016030335

PATENT

WO 2016030335

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2016030335&recNum=1&maxRec=&office=&prevFilter=&sortOption=&queryString=&tab=PCTDescription

PATENT

WO-2017031994

MACHINE TRANSLATED FROM CHINESE, BEAWARE OF FUNNY NAMES

Chinese Patent Application No. 201310275643.2 discloses a method for the synthesis of tricatadine, which uses a L-menthol ester of dihydroxyacetic acid as a raw material and undergoes condensation reaction with hydroxyacetaldehyde, and then the hydroxyl group is halogenated to obtain a halide , The halide is coupled with cytosine to obtain the coupling, and the conjugate is reduced to obtain tricatadine. However, the present inventors have found that the method requires a four-step reaction, such as condensation, halogenation, coupling and reduction, which is required to be carried out in different reaction systems. The steps are long and cumbersome, and in particular, the intermediate product is required to be separated and replaced Containers, and not suitable for amplification, it is not suitable for industrial production.

the present invention provides a process for the synthesis of a compound of formula III, wherein the synthesis reaction formula is as follows:

Example 1 Synthesis of tricatadine

The synthetic route is as follows:

Step 1: Preparation of Formula II

18.0 g of methylene chloride was added to the reaction kettle, and the mixture of the formula I was homogeneously added. After the temperature was lowered to 0 ± 3 ° C under the protection of nitrogen, 1.5 g of trimethyl iodosilane was slowly added; (V / v), and R _f = 0.5 at the point of disappearance). The reaction was carried out under nitrogen atmosphere for 2.5 ± 0.5 hours until the reaction was complete (sampling TLC test: developing solvent: petroleum ether: ethyl acetate = 4: 1 (v / v) Subsequently, the temperature of the autoclave was kept at 0 ± 3 ° C, and 3.64 g of hexamethyldisilazane and 1.15 g of N ⁴ -acetyl cytosine were slowly added dropwise . After the completion of the addition, the temperature of the control kettle was 0 ± 3 ℃, and the reaction was carried out under the protection of nitrogen for 3.5 ± 0.5 hours until the reaction was complete (sampling TLC test: developing agent: petroleum ether: ethyl acetate = 4: 1 (v / v), R _F = 0.2 points disappear).

Then, the temperature was maintained at 22 ± 3 ° C, and 10 %% (w / w) aqueous sodium thiosulfate solution was slowly added dropwise. After adding 5 g of aqueous sodium thiosulfate solution, 0.5 g of diatomaceous earth was added, hour. Filter, filter cake washed with methylene chloride 3 times, filter cake collection stand-by. The filtrate and the washing liquid were combined into the kettle, the aqueous phase and the organic phase were separated. The organic phase was washed once with 11.3 g of saturated brine. The organic phase was separated and dried overnight with anhydrous sodium sulfate to remove the water. Remove the sodium sulfate solid, the filtrate into the rotary evaporator, steaming temperature does not exceed 45 ℃, until the end of distillation. The residue obtained by steaming was transferred to a reaction vessel, 11.2 g of acetone and 18.5 g of isopropyl acetate were added, and the mixture was heated to reflux (68 ± 3 ° C) and stirred for 1 hour. Within 2.5 ± 0.5 hours, slowly cool down until the kettle temperature is 22 ± 3 ° C. The filter was filtered in a vacuum oven at about 40 ° C and dried overnight under vacuum to give a white solid (formula II).

The diatomaceous earth cake obtained by the above-mentioned filtration was transferred to a reaction vessel, heated to 27 ± 3 ° C, 18.0 g of methylene chloride was added, and the mixture was stirred and stirred for 2 hours. The filtrate was filtered and the filtrate was transferred to a rotary evaporator. The steaming temperature did not exceed 45 ° C until the distillation was completed. The crude solid obtained by steaming (the crude product of formula II) and the white solid used in the previous step were combined and transferred to a reaction kettle, and an isopropyl acetate: acetone = 3: 2 (v / v) mixed solvent was added (1 g of crude (13.3 g of isopropyl acetate + 7.9 g of acetone) was added and heated to reflux (68 ± 3 ° C), and the mixture was stirred for 1 hour. In 2.5 ± 0.5 hours, slow down to the kettle temperature of 22 ± 3 ℃. Quickly filter the filter cake with cold acetone 1.5g once. The filter cake was placed in a vacuum oven at about 40 ° C and dried overnight under vacuum to give the formula II.

Step 2: Preparation and purification of formula III

Take the type II boutique 1g added to the four bottles, add methanol 5.0g, stir the solid dispersed evenly. And 0.045 g of sodium methoxide was weighed, and the mixture was added to 0.135 g of methanol and stirred to dissolve sodium methoxide. The methanol solution of sodium methoxide was added dropwise to a four-necked flask. The incubation was carried out at 22.5 ± 2.5 ° C for 1 hour until the reaction was complete (sampling TLC: developing solvent: dichloromethane: methanol = 4: 1 (v / v) and R _f = 0.8).

After completion of the reaction, the pH of the system was adjusted to 6.5 ± 0.5 with ice acetic acid under ice bath. And then adding 200-300 mesh silica gel (available from Qingdao Ocean Chemical Plant) 10 g of sand, filling the column, the column chromatography, which was dichloromethane: methanol = 4: 1 (v / V), collecting the fraction containing tricatropa, and steaming to dryness. The steamed solid was transferred to a three-necked flask, 3.0 g of absolute ethanol was added, and the mixture was uniformly dispersed (suspended) and heated to 78 ± 2 ° C for 0.5 hour. After completion of the reflux, slowly (2.5 ± 0.5 hours) was cooled to room temperature and stirred at room temperature for 12 hours. Continue to cool down to 2.5 ± 2.5 ℃, at this temperature holding 4.5 ± 0.5 hours. Filter, filter cake with 1.0g cold ethanol washing once, thoroughly filter, the filtrate abandoned. The filter cake was transferred to a vacuum oven and dried at 38 ± 2 ° C until constant weight to obtain a purity of the formula III.

The above method can be equal to the proportion of stable amplification, for example, can be directly amplified about 60 to 180 times, that is, I feed 61.7g ~ 185.97g (other reactants equal ratio increase), after amplification of the final product (formula III) HPLC detection purity To 99.3% ~ 99.8%, the yield of 65 ~ 85%, fully meet the tricatitabine medicinal industrial needs.

PATENT

CN 104861067

PATENT

CN 105503838

PAPER

In vitro optimization of non-small cell lung cancer activity with troxacitabine, L-1,3-dioxolane-cytidine, prodrugs
Journal of medicinal chemistry (2007), 50, (9), 2249-53.

l-1,3-Dioxolane-cytidine, a potent anticancer agent against leukemia, has limited efficacy against solid tumors, perhaps due to its hydrophilicity. Herein, a library of prodrugs were synthesized to optimize in vitro antitumor activity against non-small cell lung cancer. N⁴-Substituted fatty acid amide prodrugs of 10−16 carbon chain length demonstrated significantly improved antitumor activity over l-1,3-dioxolane-cytidine. These in vitro results suggest that the in vivo therapeutic efficacy of l-1,3-dioxolane-cytidine against solid tumors may be improved with prodrug strategies.

PAPER

• Kim, Hea O.; Schinazi, Raymond F.; Shanmuganathan, Kirupathevy; Jeong, Lak S.; Beach, J. Warren; Nampalli, Satyanarayana; Cannon, Deborah L.; Chu, Chung K.
• From Journal of Medicinal Chemistry (1993), 36(5), 519-28.

PAPER

• Jin, Haolun; Tse, Allan Tse; Evans, Colleen A.; Mansour, Tarek S.; Beels, Christopher M.; Ravenscroft, Paul; Humber, David C.; Jones, Martin F.; Payne, Jeremy J.; Ramsay, Michael V. J.
• From Tetrahedron:  Asymmetry (1993), 4(2), 211-14

PAPER

• Belleau, Bernard R.; Evans, Colleen A.; Tse, H. L. Allan; Jin, Haolun; Dixit, Dilip M.; Mansour, Tarek S.
• From Tetrahedron Letters (1992), 33(46), 6949-52.

PAPER

http://pubs.acs.org/doi/pdf/10.1021/jm00089a007

J. Med. Chem. 1992,35,1987-1995 Asymmetric Synthesis of 1,3-Dioxolane-Pyrimidine Nucleosides and Their Anti-HIV Activity

References

Troxacitabine

Identifiers
IUPAC name[show]
CAS Number	145918-75-8
PubChem CID	151173
ChemSpider	399955
UNII	60KQZ0388Y
ChEMBL	CHEMBL359164
Chemical and physical data
Formula	C8H11N3O4
Molar mass	213.19 g/mol
3D model (Jmol)	Interactive image
SMILES[hide] O=C1/N=C(/N)\C=C/N1[C@H]2O[C@H](OC2)CO

//////////////TROXACITABINE, троксацитабин , تروكساسيتابين , 曲沙他滨 , Hmd-cytosineM,  NCGC00183848-01, Beta-L-Dioxolane-cytidine,   2R(-)-cis-Hmd-cytosine, (-)-ODDC

GDC 0994

GDC-0994; Ravoxertinib; 1453848-26-4; GDC0994; UNII-R6AXV96CRH; R6AXV96CRH, RG7842; RG-7842; RG 7842

CAS 1453848-26-4

1-[(1S)-1-(4-chloro-3-fluorophenyl)-2-hydroxyethyl]-4-[2-[(2-methylpyrazol-3-yl)amino]pyrimidin-4-yl]pyridin-2-one

PHASE 1

Ravoxertinib also known as GDC-0994 and RG7842, is an orally available inhibitor of extracellular signal-regulated kinase (ERK), with potential antineoplastic activity. Upon oral administration, GDC-0994 inhibits both ERK phosphorylation and activation of ERK-mediated signal transduction pathways. This prevents ERK-dependent tumor cell proliferation and survival. The mitogen-activated protein kinase (MAPK)/ERK pathway is upregulated in a variety of tumor cell types and plays a key role in tumor cell proliferation, differentiation and survival.

GDC-0994 is an ERK inhibitor invented by Array under a collaboration agreement with Genentech. Array has received certain clinical milestones and is entitled to additional potential clinical and commercial milestones and royalties on product sales under the agreement. ERK is a key protein kinase in the RAS/RAF/MEK/ERK pathway, which regulates several key cellular activities including proliferation, differentiation, migration, survival and angiogenesis. Inappropriate activation of this pathway has been shown to occur in many cancers. GDC-0994 is currently advancing in a Phase 1 trial in patients with solid tumors.

WO2013130976

• OriginatorArray BioPharma
• DeveloperGenentech
• ClassAntineoplastics; Small molecules
• Mechanism of ActionExtracellular signal-regulated MAP kinase inhibitors; Mitogen activated protein kinase 3 inhibitors; Mitogen-activated protein kinase 1 inhibitors

• Phase ISolid tumours

Most Recent Events

• 29 Nov 2016Pharmacodynamics data from a preclinical trial in Solid tumours presented at the 28^th EORTC-NCI-AACR Symposium on Molecular Targets and Cancer Therapeutics (EORTC-NCI-AACR-2016)
• 29 Nov 2016Adverse events, efficacy, pharmacokinetics and pharmacodynamics data from a phase I trial in Solid tumours presented at the 28th EORTC-NCI-AACR Symposium on Molecular Targets and Cancer Therapeutics
• 16 Jul 2016No recent reports of development identified for phase-I development in Solid-tumours(Late-stage disease, Monotherapy, Second-line therapy or greater) in USA

FREE FORM

(S)-1-(1-(4-Chloro-3-fluorophenyl)-2-hydroxyethyl)-4-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)pyridin-2(1H)-one,

GDC-0994 benzenesulfonate salt

CAS 1817728-45-2, C₂₁ H₁₈ Cl F N₆ O₂ . C₆ H₆ O₃ S

GDC-0994 as a light yellow solid,

mp 197.7 °C;

¹H NMR (600 MHz, DMSO-d₆): 9.93, (s, 1H), 8.65 (d, J = 5.2 Hz, 1H), 7.95 (d, J = 7.27 Hz, 1H), 7.63 (m, 2H), 7.62 (d, J = 1.5 Hz, 1H), 7.58 (t, J = 8.2 Hz, 1H), 7.55 (d, J = 5.2 Hz, 1H), 7.44 (dd, J = 10.6, 1.9 Hz, 1H), 7.33 (m, 3H), 7.18 (d, J = 2.0 Hz, 1H), 7.17 (d, J = 2.1 Hz, 1H), 6.90 (dd, J = 7.3, 2.1 Hz, 1H), 6.48 (d, J = 2.2 Hz, 1H), 5.99 (dd, J = 8.1, 5.5 Hz, 1H), 4.17 (dd, J = 11.9, 8.2 Hz, 1H), 4.05 (dd, J = 11.9, 5.5 Hz, 1H), 3.78 (s, 3H).

¹³C NMR (150 MHz, DMSO-d₆): 161.60, 161.14, 160.02, 159.79, 157.02 (d, J = 245 Hz), 148.0, 146.49, 139.53 (d, J = 6.0 Hz), 139.04, 136.96, 136.39, 130.66, 128.42, 127.59, 125.38, 124.99 (d, J = 3.0 Hz), 118.72 (d, J = 18.0 Hz), 117.29, 116.05 (d, J = 22.5 Hz), 109.75, 102.79, 98.77, 60.64, 58.68, 35.29.

¹⁹F NMR (282 MHz, DMSO-d₆) −115.86 (dd, J = 10.6, 7.8).

HRMS calcd for C₂₁H₁₈ClFN₆O₂ [M + H] 441.1242, found 441.1245.

PATENT

WO 2013130976

Example 39

(S)-1-(1-(4-chloro-3-fluorophenyl)-2-hydroxyethyl)-4-(2-((1-methyl-1H-pyrazol-5- yl)amino)pyrimidin-4-yl)pyridin-2(1H)-one

[00398] Step A: (S)-1-(2-(tert-Butyldimethylsiloxy)-1-(4-chloro-3-fluorophenyl)ethyl)-4-(2-(methylsulfonyl)pyrimidin-4-yl)pyridine-2(1H)-one (47 mg, 0.087 mmol), 2-methyl pyrazole-3 -amine (0.175 mmol, 2.0 equivalents) and anhydrous DMF (3.0 mL) were added to a 25 mL round bottomed flask equipped with a stirring bar. The flask was capped with a rubber septum and flushed with nitrogen. Under a blanket of nitrogen, sodium hydride (8.5 mg, 60% dispersion in mineral oil) was added in one portion. The flask was flushed with

nitrogen, capped and stirred at room temperature. The reaction progress was monitored by LCMS, and after 30 minutes, the starting material was consumed. The reaction mixture was quenched by the addition of water (0.5 mL) and ethyl acetate (15 mL). The contents of the round bottomed flask were transferred to a 125 mL separatory funnel, and the reaction flask was rinsed several times with additional ethyl acetate. Crude (S)-1-(2-((tert-butyldimethylsilyl)oxy)-1-(4-chloro-3-fluorophenyl)ethyl)-4-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)pyridin-2(1H)-one was partitioned between ethyl acetate and water (80 mL/30 mL). The ethyl acetate layer was washed once with brine, dried (MgSO₄), filtered and concentrated to give crude (S)-1-(2-((tert-butyldimethylsilyl)oxy)-1-(4-chloro-3-fluorophenyl)ethyl)-4-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)pyridin-2(1H)-one. The crude was taken directly into the deprotection step.

[00399] Step B: Crude (S)-1-(2-((tert-butyldimethylsilyl)oxy)-1-(4-chloro-3-fluorophenyl)ethyl)-4-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)pyridin-2(1H)-one (48 mg) was dissolved in ethyl acetate (4 mL) and treated dropwise slowly (over 2 minutes) with an ethyl acetate solution (1.0 mL, which had been saturated with HCl gas). The reaction stirred at room temperature for 15 minutes, after which time LCMS indicated complete consumption of the starting material. The reaction mixture was concentrated to an oily residue and purified by prep RP HPLC to yield (S)-1-(1-(4-chloro-3-fluorophenyl)-2-hydroxyethyl)-4-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)pyridin-2(1H)-one (20.8 mg, 54.6% yield) as a lyophilized powder. ¹H NMR (400 MHz, (CD₃)₂SO) δ 9.58 (s, 1H), 8.60 (d, J = 5.1 Hz, 1H), 7.91 (t, J = 9.0 Hz, 1H),7.58 (t, J = 8.1 Hz, 1H), 7.52-7.41 (m, 2H), 7.37 (d, J = 1.8 Hz, 1H), 7.14 (dd, J = 10.7,5.1 Hz 2H), 6.86 (dd, J = 7.3, 1.8 Hz, 1H), 6.27(d, J = 1.7 Hz, 1H), 5.97 (dd, J = 7.7, 5.7 Hz, 1H), 5.31(t, J = 5.2 Hz, 1H), 4.15 (m, 1H), 4.10-3.95 (m,1H), 3.69 (s, 3H); LCMS m/z 441 (M+H)+.

PAPER

Discovery of (S)-1-(1-(4-Chloro-3-fluorophenyl)-2-hydroxyethyl)-4-(2-((1-methyl-1H-pyrazol-5-yl)amino)pyrimidin-4-yl)pyridin-2(1H)-one (GDC-0994), an Extracellular Signal-Regulated Kinase 1/2 (ERK1/2) Inhibitor in Early Clinical Development

The extracellular signal-regulated kinases ERK1/2 represent an essential node within the RAS/RAF/MEK/ERK signaling cascade that is commonly activated by oncogenic mutations in BRAF or RAS or by upstream oncogenic signaling. While targeting upstream nodes with RAF and MEK inhibitors has proven effective clinically, resistance frequently develops through reactivation of the pathway. Simultaneous targeting of multiple nodes in the pathway, such as MEK and ERK, offers the prospect of enhanced efficacy as well as reduced potential for acquired resistance. Described herein is the discovery and characterization of GDC-0994 (22), an orally bioavailable small molecule inhibitor selective for ERK kinase activity.

PATENT

WO 2015154674

https://www.google.com/patents/WO2015154674A1?cl=pt

(i) i-PrMgCl， LiCl， THF； (ii) HCO₂Na， HCO₂H， H₂O， EtOH； (iii) GDH-105， morpholineethanesulfonic acid， MgCl₂， PEG6000， heptane， 1wt％ KRED-NADH-112， NAD， glucose (iv) TBSCl， DMAP， TEA， DCM， 20-25℃ ， 15h； (v) MsCl， DCM， 20-25℃ ， 3h

Scheme 1. Original Synthetic Process

Scheme 2. Improved Process To I

[0170]

Example 1

[0173]

2- ( (tert-Butyldimethylsilyl) oxy) -1- (4-chloro-3-fluorophenyl) ethyl methanesulfonate

[0174]

[0175]

Step 1: 4-bromo-1-chloro-2-fluorobenzene (64 kg) and dry toluene (170kg) were charged to the 2000 L steel reaction vessel under nitrogen. The reactor was evacuated and backfilled with N₂ for three times, and cooled to between-10 and 5 ℃ under nitrogen atmosphere. To the solution was added dropwise i-PrMgCl. LiCl (280kg, 1.3M in THF) at between-10 and 10 ℃ . The reaction was stirred for a further 15 to 30min at between-10 and 10 ℃ and then warmed to about 20 to 25 ℃ over 1h. The reaction mixture was stirred for another 6 h stir to complete the exchange. The resulting solution was cooled to between-50 and-40 ℃ . A solution of 2-chloro-N-methoxy-N-methylacetamide (44.5kg) in dry toluene (289kg) was added dropwise to the above solution at while maintaining the temperature between-50 and-30 ℃ . The reaction mixture was warmed to between 20 and 25 ℃ over 1h and then stirred for 3h to complete the reaction. The reaction was quenched by addition of 1N aq. HCl (808l g) at a temperature between-5 and 15 ℃ . The aqueous layer was separated and organic layer was filtered through a pad of diatomaceous earth. The organic layer was washed with 10％aq. NaCl solution (320kg) twice, then concentrated to about 300L to obtain 1- (4-chloro-3-fluorophenyl) -2-chloroethanone (51.8kg, 81.9％yield) as product in toluene.

[0176]

Step 2: The solution of II (51.7kg) in toluene was concentrated and solvent exchanged to EtOH to afford a suspension of II in EtOH (326kg) . A solution of HCOONa·2H₂O (54.8kg) and HCOOH (44.5kg) in water (414kg) was added at a temperature between 15 and 35 ℃ under a nitrogen atmosphere. The resulting mixture was heated to reflux and stirred for 4 to 5 h. The solution was cooled to between 20 and 30 ℃ after over 95％conversion occurred. Water (450kg) was added dropwise at between 10 and 30℃ for over 2 h. The resulting suspension was cooled to between-10 and-3 ℃ and the cooled solution stirred for 1 to 2 h. The solid was filtered and the filter cake washed with water (400 kg) to remove the residual HCOONa and HCOOH. The 1- (4-chloro-3-fluorophenyl) -2-hydroxyethanone obtained was suspended in EtOAc (41kg) and n-heptane (64kg) , then warmed to between 45 and 50 ℃ , stirred for 2h, then cooled to between-2 and 5 ℃ for over 2h and stirred at this temperature for 2h. The solids were filtered and dried in vacuo at between 40 and 50 ℃ for 12 h to afford the product as white solid (40.0kg, 99.3％purity, 84.5％yield) .

[0177]

Step 3: A 500 L reactor under nitrogen was charged with purified water (150 kg) , 4-morpholineethanesulfonic acid (0.90kg) , anhydrous MgCl₂(0.030kg) , n-heptane (37kg) , 1- (4-chloro-3-fluorophenyl) -2-hydroxyethanone (30kg) , D- (+) -glucose monohydrate (34.8kg) and PEG 6000 (30.0kg) . The pH of the solution was adjusted to between 6.5 and 7.0 with 1N aq. NaOH at between 28 and 32 ℃ . The cofactor recycling enzyme, glucose dehydrogenase GDH-105 (0.300kg) (Codexis Inc., Redwood City, CA, USA) , the cofactor nicotinamide adenine dinucleotide NAD (0.300kg) (Roche) and the oxidoreductase KRED-NADH-112 (0.300kg) (Codexis Inc., Redwood City, CA, USA) were added. The resulting suspension was stirred at between 29 and 31 ℃ for 10 to 12 h while adjusting the pH to maintain the reaction mixture pH between 6.5 and 7.0 by addition of 1N aq. NaOH (160kg) . The pH of the reaction mixture was adjusted to between 1 and 2 by addition of 49％H₂SO₄ (20kg) to quench the reaction. EtOAc (271kg) was added and the mixture was stirred at between 20 and 30 ℃for 10-15min then filtered through a pad of diatomaceous earth. The filter cake was washed with EtOAc (122kg) . The combined organic layers were separated and aqueous layer was extracted with EtOAc (150kg) . Water (237kg) was added to the combined organic layers. The pH of the mixture was adjusted to between 7.0 and 8.0 by addition of solid NaHCO₃. The organic layer was separated, concentrated and then diluted with DCM to afford (R) -1- (4-chloro-3-fluorophenyl) ethane-1, 2-diol (30.9kg, yield 100％) as product in DCM.

[0178]

Step 4: A 1000 L reactor under nitrogen was charged with (R) -1- (4-chloro-3-fluorophenyl) ethane-1, 2-diol (29.5kg) and dry DCM (390kg) . The solution was cooled to between-5 and 0 ℃ . tert-Butylchlorodimethylsilane (25.1 kg) was added in portions while maintaining the temperature between-5 and 2 ℃ . A solution of DMAP (0.95kg) and TEA (41.0kg) in dry DCM (122kg ) was added dropwise to above solution at between-5 and 2 ℃ . The reaction solution was stirred for 1 h, then warmed to between 20 and 25 ℃ and stirred for 16 h. The solution of (R) -2- ( (tert-butyldimethylsilyl) oxy) -1- (4-chloro-3-fluorophenyl) ethanol was recooled to between-10 and-5 ℃ . A solution of methanesulfonyl chloride (19.55 kg) in dry DCM (122kg) was added dropwise to the above solution of while maintaining the temperature between-10 and 0 ℃ . The reaction solution was stirred at between-10 and 0 ℃ for 20 to 30 min, and then warmed to between 0 and 5℃ for over 1h, and stirred. The reaction solution was washed with water (210kg) , followed by 5％aq. citric acid (210kg) , 2％aq. NaHCO₃ (210kg) and finally water (2 x 210kg) . The resulting DCM solution was dried (Na₂SO₄) , filtered and concentrated in vacuo below 15℃ (jacket temperature below 35℃) to afford (R) -2- ( (tert-butyldimethylsilyl) oxy) -1- (4-chloro-3-fluorophenyl) ethyl methanesulfonate (49.5kg, 83.5％yield, KarlFischer＝0.01％) as product in DCM.

[0179]

Example 2

[0180]

4- (2- (methylsulfonyl) pyrimidin-4-yl) pyridin-2 (1H) -one

[0181]

[0182]

Step 1: A 1000 L reactor was charged with 2-fluoro-4-iodopyridine (82.2kg) and dry THF (205 kg) . The reactor was evacuated and backfilled with N₂three times then cooled to between-30 and-20 ℃ . To the solution was added dropwise i-PrMgCl·LiCl (319 kg, 1.3M in THF) . The reaction was warmed to between-20 and-10℃ and stirred for 1.5 h to complete the transmetallation.

[0183]

A 2000 L reactor was charged with 4-chloro-2-methylthiopyrimidine (45.6kg) , dry THF (205kg) and [1, 3-bis (2, 6-diisopropylphenyl) imidazol-2-ylidene] (3-chloropyridyl) palladium (II) dichloride (PEPPSI^TM-IPr, 1.850kg) . The 2000 L reactor was evacuated and backfilled with N₂ three times and heated to between 55 and 57 ℃ . To the reactor was added over 0.5 to 1 h, the solution of (2-fluoropyridin-4-yl) magnesium chloride while maintaining the temperature between 50 and 62℃ . The resulting reaction mixture was stirred at between 50 and 62 ℃ for a further 2h. The reaction mixture was cooled to between 5 and 25℃ while the reaction was quenched with water (273kg) . The pH of the mixture was adjusted to 8 to 9 by adding solid citric acid monohydrate (7.3kg) . The organic layer was separated, washed with 12.5％aqNaCl (228kg) and concentrated in vacuo below 50℃ to afford 4- (2-fluoropyridin-4-yl) -2- (methylthio) pyrimidine (38.3kg, 61％yield) as product in THF.

[0184]

Step 2: The solution of 4- (2-fluoropyridin-4-yl) -2- (methylthio) pyrimidine (38.2kg) in THF was concentrated and co-evaporated with THF to remove residual water. The suspension was filtered through a pad of diatomaceous earth to remove inorganic salts. To the resulting solution in THF (510kg) was added tert-BuOK (39.7kg) in portions while maintaining the temperature between 15 and 25 ℃ . The mixture was warmed to between 20 and 25 ℃ and stirred for 5h. NaHCO₃ (14.9kg) added charged and then a citric acid solution (5kg) in THF (15kg) was added to adjust the pH to between 8 and 9. Water (230kg) was added. The mixture was filtered and the filter cake was washed with THF (100kg) . The combined THF solutions were washed with 12.5％aqueous NaCl (320kg) and concentrated to about 380L to afford a solution of 4- (2- (tert-butoxy) pyridin-4-yl) -2- (methylthio) pyrimidine in THF.

[0185]

To the THF solution cooled to between 15 and 30 ℃ was added1N H₂SO₄ aq. solution (311kg) . The mixture was stirred at this temperature for 4h. MTBE (280kg) was charged and the pH of reaction solution was adjusted to 14 with 30％aqueous NaOH (120kg) . The aqueous layer was separated and the organic phase filtered to remove inorganic salts. The obtained aqueous layer was washed with MTBE (2 x 280kg) . 2-MeTHF (1630kg) and i-PrOH (180kg) were added to the aqueous solution. The pH was then adjusted to 8 slowly with conc. HCl (19kg) . An organic layer separated and aqueous layer was extracted with 2-MeTHF (305kg) . The combined 2-MeTHF extracts were washed with water (300kg) and concentrated to about 100L. MTBE (230kg) was added and stirred at 20-30 ℃ for 0.5h. The solid was filtered and slurried in a mixture solvent of 2-MeTHF (68kg) and MTBE (230kg) . The suspension was stirred at 35-50 ℃ for 3h, and then cooled to 0 to 10 ℃ and stirred at a further 2h.

[0186]

The solid was filtered and dried in vacuo at between 50 and 62 ℃ for 20 h to afford product 4- (2- (methylthio) pyrimidin-4-yl) pyridin-2 (1H) -one as brown solid (33.55kg, 89.6％assay, 79.4％yield) .

[0187]

Example 3

[0188]

(S) -1- (2- ( (tert-Butyldimethylsilyl) oxy) -1- (4-chloro-3-fluorophenyl) ethyl) -4- (2- (methylthio) pyrimidin-4-yl) pyridin-2 (1H) -one (XI)

[0189]

[0190]

Step 1: The THF was co-evaporated from the THF solution of 4- (2- (methylthio) pyrimidin-4-yl) pyridin-2 (1H) -one (25.5kg) to remove residual water. Dry bis- (2-methoxyethyl) ether (75kg) was added. A solution of KHMDS (131kg, 1M in THF) was added dropwise while maintaining the temperature between 25 and 40 ℃ . The mixture was heated to between 75 and 80℃ and stirred for 30 to 40 min. The resulting mixture was cooled to between 20 and 30℃ under nitrogen atmosphere. A solution of (R) -2- ( (tert-butyldimethylsilyl) oxy) -1- (4-chloro-3-fluorophenyl) ethyl methanesulfonate (47.6kg) in THF (50kg) was added over 30 to 60 min while maintaining the temperature between 20 and 40℃ . The reaction solution was warmed to between 80 and 85 ℃ and stirred for 7 h. The solution was cooled to between 5 and 15 ℃ and water (155 kg) was added. The pH of the solution was adjusted to 7.5 with 30％aqueous citric acid (30 kg) . EtOAc (460kg) was added and the mixture was stirred for 20 min. The organic layer was separated and washed with 12.5％aqueous NaCl (510kg) . The combined aqueous layers were extracted with EtOAc (115kg) . The ethyl acetate layers were concentrated to about 360L to afford (S) -1- (2- ( (tert-butyldimethylsilyl) oxy) -1- (4-chloro-3-fluorophenyl) ethyl) -4- (2- (methylthio) pyrimidin-4-yl) pyridin-2 (1H) -one (44.6kg, 75.7％yield) as product in EtOAc.

[0191]

Step 2: To a solution of (S) -1- (2- ( (tert-butyldimethylsilyl) oxy) -1- (4-chloro-3-fluorophenyl) ethyl) -4- (2- (methylthio) pyrimidin-4-yl) pyridin-2 (1H) -one (44.6kg) in EtOAc (401kg, 10vol) cooled to between 5 and 10 ℃ was added in portions MCPBA (58kg) . The reaction mixture was added to a solution of NaHCO₃ (48.7kg) in water (304kg) at a temperature between10 and-20℃ . A solution of Na₂S₂O₃ (15kg) in water (150 kg) was added dropwise to consume residual MCBPA. The organic layer was separated and aqueous layer was extracted with EtOAc (130kg) . The combined organic layers were washed with water (301 kg) , concentrated and solvent exchanged to DCM to afford (S) -1- (2- ( (tert-butyldimethylsilyl) oxy) -1- (4-chloro-3-fluorophenyl) ethyl) -4- (2- (methylsulfonyl) pyrimidin- 4-yl) pyridin-2 (1H) -one (45.0kg, 94.9％yield) as product in DCM. The DCM solution was concentrated to about 100 L, filtered through a pad of SiO₂ (60kg) and eluted with an EtOAc/DCM gradient (0, 25 and 50％EtOAc) . The fractions were combined and concentrated to get the product which was re-slurried with (acetone: n-heptane＝1: 3 v/v) four times to afford the final product (31.94kg, 71％yield) .

[0192]

Example 4

[0193]

(S) -1- (1- (4-Chloro-3-fluorophenyl) -2-hydroxyethyl) -4- (2- ( (1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2 (1H) -one, benzenesulfonate salt (VIIIb)

[0194]

[0195]

Step 1: A clean 100 L cylindrical reaction vessel was charged with THF (13 kg) then (S) -1- (2- ( (tert-butyldimethylsilyl) oxy) -1- (4-chloro-3-fluorophenyl) ethyl) -4- (2- (methylsulfonyl) pyrimidin-4-yl) pyridin-2 (1H) -one (I, 5 kg) and 1-methyl-1H-pyrazol-5-amine (1.1 kg) were added sequentially with medium agitation followed by THF (18 kg) . The mixture was cooled to-35 ℃ and to the resulting thin slurry was added slowly a THF solution of LiHMDS (17.4 kg, 1.0 M) at a rate that maintained the internal temperature below-25 ℃ . After the addition was completed, the reaction was held between-35 and-25 ℃ for 20 min and monitored by HPLC. If the HPLC result indicated ≤ 98.5％conversion, additional LiHMDS (0.34 kg, 1.0 M, 0.05 mol％) was added slowly at-35 ℃ . The reaction was quenched slowly at the same temperature with H₃PO₄ solution (4.4 kg of 85％H₃PO₄and 15 kg of water) and the internal temperature was kept below 30 ℃ . The reaction was diluted with EtOAc (18 kg) and the phases separated, the organic layer was washed with H₃PO₄ solution (1.1 kg of 85％H₃PO₄ and 12 kg of water) followed by a second H₃PO₄wash (0.55 kg of 85％H₃PO₄and 12 kg of water) . If 1-methyl-1H-pyrazol-5-remained, the organic layer was washed again with H₃PO₄ solution (0.55 kg of 85％H₃PO₄ and 12 kg of water) . Finally the organic layer was washed sequentially with water (20 kg) and a NaCl and NaHCO₃ solution (2 kg of NaCl, 0.35 kg of NaHCO₃and 10 kg of water) . After the phase separation, residue water in organic solution was removed through an azeotropic distillation with EtOAc to ≤ 0.5％ (by KF) and then solution was concentrated to 20-30 L under a vacuum below 50 ℃ . The solvent was then swapped to MeOH using 35 kg of MeOH and then concentrated to between 20 and 30 L for the next step.

[0196]

Step 2: To the methanolic (S) -1- (2- ( (tert-butyldimethylsilyl) oxy) -1- (4-chloro-3-fluorophenyl) ethyl) -4- (2- ( (1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2 (1H) -one (IX) solution in MeOH was added HCl (10.7 kg, 1.25 M in MeOH) at RT. It was slightly exothermic. After the addition was completed, the reaction was heated to 45 ℃ . If the reaction was incomplete after 14 to 16 h, additional HCl (1 kg, 1.25 M in MeOH) was added and agitation at 45 ℃ was continued for 2 h. The reaction was equipped with a distillation setup with acid scrubber. The reaction was concentrated to between 20 and 30 L under a vacuum below 50 ℃ . To the resulting solution was added MeOH (35 kg) and the reaction was concentrated to 20 to 30 L again under a vacuum below 50 ℃ . The solvent was then switched to EtOAc using 40 kg of EtOAc. The solvent ratio was monitored by Headspace GC and the solvent swap continued until it was less than 1/5. The solution was concentrated to between 20 and 30 L under a vacuum below 50 ℃ . After the solution was cooled below 30 ℃ , aqueous NaHCO₃ (1.2 kg of NaHCO₃ and 20 kg of water) was added slowly with a medium agitation and followed by EtOAc (40 kg) . The organic layer was washed with water (2 x 10 kg) then concentrated to 20-30 L under a vacuum below 50 ℃ . The solvent was then switched to MEK using 35 kg of MEK. The residue MeOH was monitored by Headspace GC and the solvent swap continued until the MeOH was < 0.3％. The solution containing (S) -1- (1- (4-chloro-3-fluorophenyl) -2-hydroxyethyl) -4- (2- ( (1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2 (1H) -one (VIII) was concentrated to 20 to 30 L under a vacuum below 50 ℃ for the next step.

[0197]

Step 3: The solution of VIII in MEK was transferred to a second 100 L cylindrical reaction vessel through a 1μm line filter. In a separate container was prepared benezenesulfonic acid solution (1.3 kg of benzenesulfonic acid, 1.4 kg of water and 4.4 kg of MEK) . The filtered VIII solution was heated to 75 ℃ and to the resulting solution was added 0.7 kg of the benzenesulfonic acid solution through a 1μm line filter. The clear solution was seeded with crystalline benzenesulfonic acid salt of VIII (0.425 kg) as a slurry in MEK (0.025 kg of VIIIb crystalline seed and 0.4 kg of MEK) which produced a thin slurry. The remaining benzenesulfonic acid solution was then added through a 1μm line filter in 2 h. After addition, the slurry was heated at 75 ℃ for additional 1 h and then cooled to 18 ℃ in a minimum of 3 h. The resulting thick slurry was agitated at 20 ℃ for 14 to 16 h. The solid was filtered using an Aurora dryer. The mother liquor was assayed by HPLC (about . 3％loss) . The solid was then washed with 1μm line filtered 15.8 kg of MEK and water solution (0.8 kg of water and 15 kg of MEK) and followed by 1μm line filtered 30 kg of MEK. Washes were assayed by HPLC (<1％loss) . The wet cake was dried under a vacuum and a nitrogen sweep at a jacket temperature of 45 ℃ for a minimum 12 h to afford the benzenesulfonic acid salt of VIII, which is labeled VIIIb.

[0198]

Additional Examples

[0199]

Step 1:

[0200]

[0201]

To a clean 100 L cylindrical reaction vessel was charged 13 kg of THF first. With a medium agitation, 5.0 kg of I and 1.1 kg of 1-methyl-1H-pyrazol-5-amine was charged sequentially and followed by the rest of THF (18 kg) . At-35 ℃ to the resulting thin slurry was added 17.4 kg of LiHMDS (1.0 mol/L) in THF slowly and the internal temperature was remained below-25 ℃ . After addition, the reaction was held between-35 and-25 ℃ for 20 min. The reaction was monitored by HPLC. If the HPLC result indicated ≤ 98.5％conversion, additional 0.34 kg (0.05 mol％) of LiHMDS (1.0 mol/L) in THF was charged slowly at-35 ℃ . Otherwise, the reaction was quenched at the same temperature with 19.4 kg of H₃PO₄ solution (4.4 kg of 85％H₃PO₄and 15 kg of water) slowly and the internal temperature was remained below 30 ℃ . The reaction was diluted with 18 kg of EtOAc. After the phase separation, the organic layer was washed with 13.1 kg of H₃PO₄ solution (1.1 kg of 85％H₃PO₄ and 12 kg of water) and then with 12.6 kg of H₃PO₄solution (0.55 kg of 85％H₃PO₄ and 12 kg of water) . The organic layer was assayed for the 1-methyl-1H-pyrazol-5-amine level by HPLC. If the HPLC result indicated ≥ 20 μg/mL of 1-methyl-1H-pyrazol-5-amine, the organic layer needed an additional wash with 12.6 kg of H₃PO₄ solution (0.55 kg of 85％H₃PO₄ and 12 kg of water) . Otherwise, the organic layer was washed with 20 kg of water. The organic layer was assayed again for the 1-methyl-1H-pyrazol-5-amine level. If the HPLC result indicated ≥ 2 μg/mL of 1-methyl-1H-pyrazol-5-amine, the organic layer needed an additional wash with 20 kg of water. Otherwise, the organic layer was washed with 12.4 kg of NaCl and NaHCO₃ solution (2 kg of NaCl, 0.35 kg of NaHCO₃ and 10 kg of water) . After the phase separation, residue water in organic solution was removed through an azeotropic distillation with EtOAc to ≤ 0.5％ (by KF) and then the solution was concentrated to 20 to 30 L under a vacuum below 50 ℃ . The solvent was then swapped to MeOH using 35 kg of MeOH and then concentrated to 20 to 30 L for the next step.

[0202]

Step 2:

[0203]

[0204]

To the IX solution in MeOH from the last step was charged 10.7 kg of HCl (1.25 M in MeOH) at the ambient temperature. It was observed slightly exothermic. After addition, the reaction was heated to 45 ℃ . After 14-16 h, the reaction was monitored by HPLC. If the HPLC result indicated the conversion was ≤ 98％, an additional 1 kg of HCl (1.25 M in MeOH) was charged and the reaction was agitated at 45 ℃ for additional 2 h. Otherwise, the reaction was equipped with a distillation setup with acid scrubber. The reaction was concentrated to 20 to 30 L under a vacuum below 50 ℃ . To the resulting solution was charged 35 kg of MeOH and the reaction was concentrated to 20 to 30 L again under a vacuum below 50 ℃ . The solvent was then switched to EtOAc using 40 kg of EtOAc. The solvent ratio was monitored by Headspace GC. If the ratio of MeOH/EtOAc was greater than 1/5, the solvent swap should be continued. Otherwise, the solution was concentrated to 20 to 30 L under a vacuum below 50 ℃ . After the solution was cooled below 30 ℃ , 21.2 kg of NaHCO₃ solution (1.2 kg of NaHCO₃ and 20 kg of water) was charged slowly with a medium agitation and followed by 40 kg of EtOAc. After the phase separation, the organic layer was washed with 2 X 10 kg of water. The organic layer was concentrated to 20 to 30 L under a vacuum below 50 ℃ . The solvent was then switched to MEK using 35 kg of MEK. The residue MeOH was monitored by Headspace GC. If the level of MeOH was ≥ 0.3％, the solvent swap should be continued. Otherwise, the solution was concentrated to 20 to 30 L under a vacuum below 50 ℃ for the next step.

[0205]

Step 3:

[0206]

[0207]

The VIII solution in MEK from the last step was transferred to a second 100 L cylindrical reaction vessel through a 3 μm line filter. In a separated container was prepared 7.1 kg of benzenesulfonic acid solution (1.3 kg of benzenesulfonic acid, 1.4 kg of water and 4.4 kg of MEK) . The filtered G02584994 solution was heated to 75 ℃ and to the resulting solution was charged 0.7 kg of benzenesulfonic acid solution (10％) through a 3 μm line filter. To the clear solution was charged 0.425 kg of VIIIb crystalline seed slurry in MEK (0.025 kg of VIIIb crystalline seed and 0.4 kg of MEK) . This resulted in a thin slurry. The rest of benzenesulfonic acid solution was then charged through a 3 μm line filter in 2 h. After addition, the slurry was heated at 75 ℃ for additional 1 h and then cooled to 20 ℃ in a minimum of 3 h. The resulting thick slurry was agitated at 20 ℃ for 14-16 h. Solid was filtered using a filter dryer. Mother liquor was assayed by HPLC (about 3％loss) . Solid was then washed with 3 μm line filtered 15.8 kg of MEK and water solution (0.8 kg of water and 15 kg of MEK) and followed by 3 μm line filtered 30 kg of MEK. Washes were assayed by HPLC (<1％loss) . The wet cake was dried under a vacuum and the nitrogen sweep at a jacket temperature of 45 ℃ for a minimum 12 h.

[0208]

Recrystallization

[0209]

[0210]

To a clean 100 L cylindrical reaction vessel was charged 16 kg of EtOH first. With a medium agitation, 3.5 kg of VIIIb was charged and then followed by the rest of EtOH (8.5 kg) . The thick slurry was heated to 78 ℃ and water (～1.1 kg) was charge until a clear solution was obtained. The hot solution was filtered through a 3 μm line filter to a second clean 100 L cylindrical reaction vessel. The temperature dropped to 55-60 ℃ and the solution remained clear. To the resulting solution was charged with 0.298 kg of VIIIb crystalline seed slurry in EtOH (0.018 kg of VIIIb crystalline seed and 0.28 kg of EtOH) . The thick slurry was concentrated to 20 to 30 L at 60 ℃ under a vacuum and then cooled 20 ℃ in 3 h. The resulting slurry was agitated at 20 ℃ for 14 to 16 h. Solid was filtered using a filter dryer. The mother liquor was assayed by HPLC (about 10％loss) . Solid was then washed with 3 μm line filtered 11.1 kg of EtOH and water solution (0.56 kg of water and 11 kg of EtOH) and followed by 3 μm line filtered 21 kg of MEK. Washes were assayed by HPLC (3％loss) . The wet cake was dried under a vacuum and the nitrogen sweep at a jacket temperature of 45 ℃ for a minimum 12 h.

[0211]

An additional synthetic process is set forth below.

[0212]

Step 1:

[0213]

[0214]

To a clean 100 L cylindrical reaction vessel was charged 18 kg of THF first. With a medium agitation, 4.2 kg of I and 0.91 kg of 1-methyl-1H-pyrazol-5-amine was charged sequentially and followed by the rest of THF (21 kg) . At-40 ℃ to the resulting thin slurry was added 14.9 kg of LiHMDS (1.0 mol/L) in THF slowly and the internal temperature was remained below-30 ℃ . After addition, the reaction was held between-35 and-40 ℃ for 20 min. The reaction was monitored by HPLC. The HPLC result indicated 99.1％conversion. The reaction was quenched at the same temperature with 16.7 kg of H₃PO₄ solution (3.7 kg of 85％H₃PO₄ and 13 kg of water) slowly and the internal temperature was remained below 30 ℃ . The reaction was diluted with 17 kg of EtOAc. After the phase separation, the organic layer was washed with 13.1 kg of H₃PO₄ solution (1.1 kg of 85％H₃PO₄ and 12 kg of water) and then with 10.5 kg of H₃PO₄ solution (0.46 kg of 85％H₃PO₄ and 10 kg of water) . The organic layer was assayed for the 1-methyl-1H-pyrazol-5-amine level by HPLC. The HPLC result indicated 2 μg/mL of 1-methyl-1H-pyrazol-5-amine. The organic layer was washed with 15.8 kg of NaCl solution (0.3 kg of NaCl and 15.5 kg of water) . The organic layer was assayed again for the G02586778 level. The HPLC result indicated 0.5 μg/mL of 1-methyl-1H-pyrazol-5-amine. The organic layer was washed with 10.3 kg of NaCl and NaHCO₃ solution (1.7 kg of NaCl, 0.6 kg of NaHCO₃and 8 kg of water) . After the phase separation, residue water in organic solution was removed through an azeotropic distillation with EtOAc to ≤ 0.5％ (by KF) and then the solution was concentrated to 20 to 30 L under a vacuum below 50 ℃ . The solvent was then swapped to MeOH using 30 kg of MeOH and then concentrated to 20 to 30 L for the next step.

[0215]

Step 2:

[0216]

[0217]

To the IX solution in MeOH from the last step was charged 9.0 kg of HCl (1.25 M in MeOH) at the ambient temperature. It was observed slightly exothermic. After addition, the reaction was heated to 45 ℃ . After 16 h, the reaction was monitored by HPLC. The HPLC result indicated the conversion was 99.4％. The reaction was equipped with a distillation setup. The reaction was concentrated to 20 L under a vacuum below 50 ℃ . To the resulting solution was charged 35 kg of MeOH and the reaction was concentrated to 20 L again under a vacuum below 50 ℃ . The solvent was then switched to EtOAc using 40 kg of EtOAc. The solvent ratio was monitored by Headspace GC. If the ratio of MeOH/EtOAc was greater than 1/5, the solvent swap should be continued. Otherwise, the solution was concentrated to 20 L under a vacuum below 50 ℃ . After the solution was cooled below 30 ℃ , 18 kg of NaHCO₃ solution (1 kg of NaHCO₃ and 17 kg of water) was charged slowly with a medium agitation and followed by 34 kg of EtOAc. After the phase separation, the organic layer was washed with 2 X 8 kg of water. The organic layer was concentrated to 20 L under a vacuum below 50 ℃ . The solvent was then switched to MEK using 35 kg of MEK. The residue MeOH was monitored by Headspace GC. If the level of MeOH was ≥ 0.3％, the solvent swap should be continued. Otherwise, the solution was concentrated to 20 L under a vacuum below 50 ℃ for the next step.

[0218]

Step 3:

The VIII solution in MEK from the last step was transferred to a second 100 L cylindrical reaction vessel through a 1 μm polish filter. In a separated container was prepared 6.0 kg of benzenesulfonic acid solution (1.1 kg of benzenesulfonic acid, 1.2 kg of water and 3.7 kg of MEK) . The filtered solution was heated to 75 ℃ and to the resulting solution was charged 0.6 kg of benzenesulfonic acid solution (10％) through a 1 μm line filter. To the clear solution was charged 0.36 kg of VIIIb crystalline seed slurry in MEK (0.021 kg of VIIIb crystalline seed and 0.34 kg of MEK) . This resulted in a thin slurry. The rest of benzenesulfonic acid solution was then charged through a 1 μm line filter in 2 h. After addition, the slurry was heated at 75 ℃ for additional 1 h and then cooled to 18 ℃ in a minimum of 3 h. The resulting thick slurry was agitated at 18 ℃ for 14-16 h. Solid was filtered using an Aurora dryer. Solid was then washed with 1 μm line filtered 8.15 kg of MEK and water solution (0.35 kg of water and 7.8 kg of MEK) and followed by 1 μm line filtered 12 kg of MEK.

Recrystallization

To a clean 100 L cylindrical reaction vessel was charged 21 kg of EtOH first. With a medium agitation, 3.5 kg of VIIIb was charged and then followed by the rest of EtOH (9 kg) . The thick slurry was heated to 78 ℃ and water (1.2 kg) was charge until a clear solution was obtained. The hot solution was filtered through a 1 μm line filter to a second clean 100 L cylindrical reaction vessel. The temperature dropped to 69 ℃ and the solution remained clear. To the resulting solution was charged with 0.37 kg of VIIIb crystalline seed slurry in EtOH (0.018 kg of VIIIb crystalline seed and 0.35 kg of EtOH) . The thin slurry was concentrated to 20 L at 60-70 ℃ under a vacuum and then cooled 18 ℃ in 3 h. The resulting slurry was agitated at 18 ℃ for 14-16 h. Solid was filtered using a filter dryer. Solid was then washed with 1 μm line filtered 8.6 kg of EtOH and water solution (0.4 kg of water and 8.2 kg of EtOH) . The solution was introduced in two equal portions. The solid was then washed by 1 μm line filtered 6.7 kg of MEK. The wet cake was dried under a vacuum and the nitrogen sweep at a jacket temperature of 35-40 ℃ for a minimum 12 h.

Alternative Synthetic Route (Steps 1 to 10 below)

Step 1:

[0227]

[0228]

Procedure:

[0229]

1. Charge compound 1 and MeBrPPh₃ to a four-necked jacketed flask with a paddle stirrer under N₂

[0230]

2. Charge THF (5.0V., KF<0.02％) to the flask (Note: V is the volume of solution to mass of limited reagent or L/Kg)

[0231]

3. Stir the suspension at 0 ℃

[0232]

4. Add the NaH (60％suspended in mineral oil) portionwise to the flask at 0 ℃

[0233]

5. Stir at 0 ℃ for 30min

[0234]

6. Heat to 30 ℃ and stir for 6 hrs

[0235]

7. Cool to 0 ℃

[0236]

8. Charge PE (petroleum ether) (5.0V. ) to the flask

[0237]

9. Add the crystal seed of TPPO (triphenylphospine oxide) (1 to about 5％wt of total TPPO) to the flask

[0238]

10. Stir at-10 ℃ for 2hrs

[0239]

11. Filter, and wash the cake with PE (5.0V. )

[0240]

12. Concentrate the filtrate to dryness

[0241]

13. Purification of the product by distillation under reduced pressure affords 2 as colorless oil

[0242]

Step 2:

[0243]

[0244]

Procedure:

[0245]

1. Add (DHQD) 2PHAL, Na₂CO₃, K₂Fe (CN) ₆, K₂OsO₂ (OH) ₄ into a flask under N₂ (Ad-mix beta, Aldrich, St. Louis, MO) .

[0246]

2. Cool to 0 ℃

[0247]

3. Add tBuOH (5V) and H₂O (5V)

[0248]

4. Add 2

[0249]

5. Stir the mixture at 0 ℃ for 6h

[0250]

6. Cool to 0 ℃

[0251]

7. Add Na₂SO₃ to quench the reaction

[0252]

8. Stir at 0 ℃ for 2h

[0253]

9. Filter and wash the cake with EA (ethyl acetate)

[0254]

10. Separate the organic layer

[0255]

11. Filter and concentrate to dryness

[0256]

Step 3:

[0257]

[0258]

Procedure:

[0259]

1. Add IV (1 eq. ) and DCM (5V) to a flask under N₂

[0260]

2. Cool to 0 ℃

[0261]

3. Add DMAP (0.1 eq. ) , then TEA (1.5 eq. )

[0262]

4. Add TBSCl (1.05 eq. ) dropwise at 0 ℃

[0263]

5. Stir the mixture at 0 ℃ for 1h

[0264]

6. Add water to quench the reaction

[0265]

7. Separate the layers

[0266]

8. Dry the organic layer over Na₂SO₄

[0267]

9. Filter

[0268]

10. Concentrate the filtrate to dryness

[0269]

11. Use for next step directly

[0270]

Step 4:

[0271]

[0272]

Procedure:

[0273]

1. Add V (1.0 eq. ) and DCM (5V) into a flask under N₂.

[0274]

2. Cool to 0 ℃

[0275]

3. Add TEA (1.51 eq. )

[0276]

4. Add MsCl (1.05 eq. ) dropwise at 0 ℃

[0277]

5. Stir the mixture at rt for 1h

[0278]

6. Add DCM to dilute the mixture for better stirring

[0279]

7. Add water to quench the reaction

[0280]

8. Separate the layers

[0281]

9. Wash the organic layer with NaHCO₃

[0282]

10. Dry over Na₂SO₄

[0283]

11. Filter and concentrate the filtrate to dryness

[0284]

12. Used for next step directly

[0285]

Step 5:

[0286]

[0287]

Procedure:

[0288]

1. Add VII (1eq. ) and DGME (20V) into flask under N₂

[0289]

2. Cool to 0 ℃

[0290]

3. Add KHMDS (1M in THF, 1 eq. )

[0291]

4. Add VI (1.2-1.5 eq. ) in DGME solution

[0292]

5. Stir at 0 ℃ for 5min

[0293]

6. Heat to reflux (jacket 120 ℃) and stir for over 4h

[0294]

7. Cool down

[0295]

8. Quench with water and extraction with MTBE

[0296]

9. Wash with 20％NaCl

[0297]

10. Dry over Na₂SO₄

[0298]

11. Concentrate to dryness and use to next step directly

[0299]

Step 6:

[0300]

[0301]

Procedure:

[0302]

1. Charge XI (1eq. ) , DCM (8V) into flask under N₂

[0303]

2. Add mCPBA by portions

[0304]

3. Stir at room temperature for 2h

[0305]

4. Add 7％NaHCO₃ aq. to wash

[0306]

5. Quench with Na₂S₂O₄ aq.

[0307]

6. Wash with 20％NaCl aq.

[0308]

7. Dry over Na₂SO₄

[0309]

8. Filter and concentrate to dryness

[0310]

9. Slurry the result in MTBE (3V) to afford I

[0311]

Step 7:

[0312]

[0313]

Procedure:

[0314]

1. Add I (1eq. ) , 1-methyl-1H-pyrazol-5-amine (4 eq. ) , Cs₂CO₃, DMF (4V) into a flask under N₂

[0315]

2. Stir at room temperature for 3h

[0316]

3. Work-up to afford product.

[0317]

Step 8:

[0318]

[0319]

Procedure:

[0320]

1. IX was dissolved in MeOH

[0321]

2. HCl (1.25 M in MeOH) was charged at the ambient temperature.

[0322]

3. After addition, the reaction was heated to 45 ℃ for 16 h.

[0323]

4. The reaction was cooled to rt and quenched with aqueous NaHCO₃ and diluted with EtOAc

[0324]

5. After the phase separation, the organic layer was washed with water. The organic layer was concentrated to afford the crude VIII

[0325]

Step 9:

[0326]

[0327]

Procedure:

[0328]

1. Charge compound 6-2, XIII, Pd-catalyst and sodium bicarbonate to a four-necked jacketed flask with paddle stirrer under N₂

[0329]

2. Charge water and 1, 4-dioxane (5.0V., KF<0.02％) to the flask

[0330]

3. Stir the suspension at 85 ℃ for 16hrs

[0331]

4. Filter through the silica-gel (2.0 X) and diatomaceous earth (0.5X)

[0332]

5. Remove the 1, 4-dioxane by distillation under a vacuum

[0333]

6. Partition between water (2.0V) and EtOAc (5.0V)

[0334]

7. Separate the organic phase and concentrate

[0335]

8. Purify by re-crystallization from PE and EtOAc

[0336]

Step 10:

[0337]

[0338]

Procedure:

[0339]

● Add X into a flask

[0340]

● Add 2M HCl (10-15V)

[0341]

● Heat to 100 ℃ and stir for 3h

[0342]

● Cool down

[0343]

● Neutralize pH to 7 to 8 with 30％NaOH aq.

[0344]

● Extract with THF

[0345]

● Wash with 20％NaCl aq.

[0346]

● Dry over Na₂SO₄

[0347]

● Filter and concentrate to dryness

[0348]

Synthesis of Crystalline (S) -1- (1- (4-chloro-3-fluorophenyl) -2-hydroxyethyl) -4- (2- ( (1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2 (1H) -one benzenesulfonate salt

[0349]

(S) -1- (1- (4-chloro-3-fluorophenyl) -2-hydroxyethyl) -4- (2- ( (1-methyl-1H-pyrazol-5-yl) amino) pyrimidin-4-yl) pyridin-2 (1H) -one (21.1 mg, 0.048 mmol) was dissolved in MEK (0.5 mL) . Benzenesulfonic acid (Fluka, 98％, 7.8 mg, 0.049 mmol) was dissolved in MEK (0.5 mL) and the resulting solution added drop wise to the free base solution with stirring. Precipitation occurred and the precipitate slowly dissolved as more benzenesulfonic acid solution was added. A small amount of sticky solid remained on the bottom of the vial. The vial contents were sonicated for 10 minutes during which further precipitation occurred. The solid was isolated after centrifugation and vacuum dried at 40 ℃ using house vacuum.

A process for the preparation of a compound of formula VIII, the process comprising the steps of:

(a) contacting 4-bromo-1-chloro-2-fluorobenzene with a metallating agent in an aprotic organic solvent to afford an organomagnesium compound, which is reacted with 2-chloro-N-methoxy-N-methylacetamide to afford 2-chloro-1- (4-chloro-3-fluorophenyl) ethanone (II) ；

(b) contacting II with sodium formate and formic acid in aqueous ethanol to afford 1- (4-chloro-3-fluorophenyl) -2-hydroxyethanone (III)

(c) contacting III with a ketoreductase to afford (R) -1- (4-chloro-3-fluorophenyl) ethane-1, 2-diol (IV) ；

(d) contacting IV with a silyl chloride (R^a) ₃SiCl and at least one base in a non-polar aprotic solvent to afford (V) , and subsequently adding sulfonylchloride R^bS (O) ₂Cl to afford VI, wherein R^a is independently in each occurrence C_1-6 alkyl or phenyl and R^b is selected from C_1-4 alkyl or phenyl, optionally substituted with 1 to 3 groups independently selected from C_1-3 alkyl, halogen, nitro, cyano, or C_1-3 alkoxy；

(e) contacting 4- (2- (methylsulfonyl) pyrimidin-4-yl) pyridin-2 (1H) -one (VII) with a strong base in an organic solvent and subsequently adding VI to afford XI；

(f) treating XI with an oxidizing agent to afford I；

(g) treating 1-methyl-1H-pyrazol-5-amine with a strong base in an aprotic solvent at reduced temperature and adding the compound of formula I to afford IX； and,

(h) contacting IX with a de-silylating agent to afford VIII.

PAPER

Development of a Practical Synthesis of ERK Inhibitor GDC-0994

Xin Linghu^*† , Nicholas Wong^†, Hans Iding^‡, Vera Jost^‡, Haiming Zhang^†, Stefan G. Koenig^†, C. Gregory Sowell^†, and Francis Gosselin^†

^† Small Molecule Process Chemistry, Genentech, Inc., 1 DNA Way, South San Francisco, California 94080, United States

^‡ Process Research, F. Hoffmann-La Roche AG, Grenzacherstrasse 124, CH-4070 Basel, Switzerland

Org. Process Res. Dev., Article ASAP

DOI: 10.1021/acs.oprd.7b00006

http://pubs.acs.org/doi/abs/10.1021/acs.oprd.7b00006

The process development of a synthetic route to manufacture ERK inhibitor GDC-0994 on multikilogram scale is reported herein. The API was prepared as the corresponding benzenesulfonate salt in 7 steps and 41% overall yield. The synthetic route features a biocatalytic asymmetric ketone reduction, a regioselective pyridone S_N2 reaction, and a safe and scalable tungstate-catalyzed sulfide oxidation. The end-game process involves a telescoped S_NAr/desilylation/benzenesulfonate salt formation sequence. Finally, the development of the API crystallization allowed purging of process-related impurities, obtaining >99.5A% HPLC and >99% ee of the target molecule.

///////////GDC 0994, Ravoxertinib, 1453848-26-4, GDC0994, UNII-R6AXV96CRH, R6AXV96CRH, RG7842, RG-7842, RG 7842, PHASE 1

CN1C(=CC=N1)NC2=NC=CC(=N2)C3=CC(=O)N(C=C3)C(CO)C4=CC(=C(C=C4)Cl)F

• Approved based on a first-line Phase III trial that met its primary endpoint of progression-free survival (PFS) at interim analysis due to superior efficacy compared to letrozole alone[1]
• At this interim analysis, Kisqali plus letrozole reduced risk of disease progression or death by 44% over letrozole alone, and demonstrated tumor burden reduction with a 53% overall response rate[1]
• Kisqali plus letrozole showed treatment benefit across all patient subgroups regardless of disease burden or tumor location[1]
• At a subsequent analysis with additional follow-up and progression events, a median PFS of 25.3 months for Kisqali plus letrozole and 16.0 months for letrozole alone was observed[2]

Basel, March 13, 2017 – The US Food and Drug Administration (FDA) has approved Kisqali^®(ribociclib, formerly known as LEE011) in combination with an aromatase inhibitor as initial endocrine-based therapy for treatment of postmenopausal women with hormone receptor positive, human epidermal growth factor receptor-2 negative (HR+/HER2-) advanced or metastatic breast cancer.

Kisqali is a CDK4/6 inhibitor approved based on a first-line Phase III trial that met its primary endpoint early, demonstrating statistically significant improvement in progression-free survival (PFS) compared to letrozole alone at the first pre-planned interim analysis[1]. Kisqali was reviewed and approved under the FDA Breakthrough Therapy designation and Priority Review programs.

“Kisqali is emblematic of the innovation that Novartis continues to bring forward for people with HR+/HER2- metastatic breast cancer,” said Bruno Strigini, CEO, Novartis Oncology. “We at Novartis are proud of the comprehensive clinical program for Kisqali that has led to today’s approval and the new hope this medicine represents for patients and their families.”

The FDA approval is based on the superior efficacy and demonstrated safety of Kisqali plus letrozole versus letrozole alone in the pivotal Phase III MONALEESA-2 trial. The trial, which enrolled 668 postmenopausal women with HR+/HER2- advanced or metastatic breast cancer who received no prior systemic therapy for their advanced breast cancer, showed that Kisqali plus an aromatase inhibitor, letrozole, reduced the risk of progression or death by 44 percent over letrozole alone (median PFS not reached (95% CI: 19.3 months-not reached) vs. 14.7 months (95% CI: 13.0-16.5 months); HR=0.556 (95% CI: 0.429-0.720); p<0.0001)[1].

More than half of patients taking Kisqali plus letrozole remained alive and progression free at the time of interim analysis, therefore median PFS could not be determined[1]. At a subsequent analysis with additional 11-month follow-up and progression events, a median PFS of 25.3 months for Kisqali plus letrozole and 16.0 months for letrozole alone was observed[2]. Overall survival data is not yet mature and will be available at a later date.

“In the MONALEESA-2 trial, ribociclib plus letrozole reduced the risk of disease progression or death by 44 percent over letrozole alone, and more than half of patients (53%) with measurable disease taking ribociclib plus letrozole experienced a tumor burden reduction of at least 30 percent. This is a significant result for women with this serious form of breast cancer,” said Gabriel N. Hortobagyi, MD, Professor of Medicine, Department of Breast Medical Oncology, The University of Texas MD Anderson Cancer Center and MONALEESA-2 Principal Investigator. “These results affirm that combination therapy with a CDK4/6 inhibitor like ribociclib and an aromatase inhibitor should be a new standard of care for initial treatment of HR+ advanced breast cancer.”

Kisqali is taken with or without food as a once-daily oral dose of 600 mg (three 200 mg tablets) for three weeks, followed by one week off treatment. Kisqali is taken in combination with four weeks of any aromatase inhibitor[1].

Breast cancer is the second most common cancer in American women[3]. The American Cancer Society estimates more than 250,000 women will be diagnosed with invasive breast cancer in 2017[3]. Up to one-third of patients with early-stage breast cancer will subsequently develop metastatic disease[4].

Novartis is committed to providing patients with access to medicines, as well as resources and support to address a range of needs. The Kisqali patient support program is available to help guide eligible patients through the various aspects of getting started on treatment, from providing educational information to helping them understand their insurance coverage and identify potential financial assistance options. For more information, patients and healthcare professionals can call 1-800-282-7630.

The full prescribing information for Kisqali can be found at https://www.pharma.us.novartis.com/sites/www.pharma.us.novartis.com/files/kisqali.pdf(link is external).

About Kisqali^® (ribociclib)
Kisqali (ribociclib) is a selective cyclin-dependent kinase inhibitor, a class of drugs that help slow the progression of cancer by inhibiting two proteins called cyclin-dependent kinase 4 and 6 (CDK4/6). These proteins, when over-activated, can enable cancer cells to grow and divide too quickly. Targeting CDK4/6 with enhanced precision may play a role in ensuring that cancer cells do not continue to replicate uncontrollably.

Kisqali was developed by the Novartis Institutes for BioMedical Research (NIBR) under a research collaboration with Astex Pharmaceuticals.

About the MONALEESA Clinical Trial Program
Novartis is continuing to assess Kisqali through the robust MONALEESA clinical trial program, which includes two additional Phase III trials, MONALEESA-3 and MONALEESA-7, that are evaluating Kisqali in multiple endocrine therapy combinations across a broad range of patients, including premenopausal women. MONALEESA-3 is evaluating Kisqali in combination with fulvestrant compared to fulvestrant alone in postmenopausal women with HR+/HER2- advanced breast cancer who have received no or a maximum of one prior endocrine therapy. MONALEESA-7 is investigating Kisqali in combination with endocrine therapy and goserelin compared to endocrine therapy and goserelin alone in premenopausal women with HR+/HER2- advanced breast cancer who have not previously received endocrine therapy.

About Novartis in Advanced Breast Cancer
For more than 25 years, Novartis has been at the forefront of driving scientific advancements for breast cancer patients and improving clinical practice in collaboration with the global community. With one of the most diverse breast cancer pipelines and the largest number of breast cancer compounds in development, Novartis leads the industry in discovery of new therapies and combinations, especially in HR+ advanced breast cancer, the most common form of the disease.

Kisqali^® (ribociclib) Important Safety Information
Kisqali^® (ribociclib) can cause a heart problem known as QT prolongation. This condition can cause an abnormal heartbeat and may lead to death. Patients should tell their healthcare provider right away if they have a change in their heartbeat (a fast or irregular heartbeat), or if they feel dizzy or faint. Kisqali can cause serious liver problems. Patients should tell their healthcare provider right away if they get any of the following signs and symptoms of liver problems: yellowing of the skin or the whites of the eyes (jaundice), dark or brown (tea-colored) urine, feeling very tired, loss of appetite, pain on the upper right side of the stomach area (abdomen), and bleeding or bruising more easily than normal. Low white blood cell counts are very common when taking Kisqali and may result in infections that may be severe. Patients should tell their healthcare provider right away if they have signs and symptoms of low white blood cell counts or infections such as fever and chills. Before taking Kisqali, patients should tell their healthcare provider if they are pregnant, or plan to become pregnant as Kisqali can harm an unborn baby. Females who are able to become pregnant and who take Kisqali should use effective birth control during treatment and for at least 3 weeks after the last dose of Kisqali. Do not breastfeed during treatment with Kisqali and for at least 3 weeks after the last dose of Kisqali. Patients should tell their healthcare provider about all of the medicines they take, including prescription and over-the-counter medicines, vitamins, and herbal supplements since they may interact with Kisqali. Patients should avoid pomegranate or pomegranate juice, and grapefruit or grapefruit juice while taking Kisqali. The most common side effects (incidence >=20%) of Kisqali when used with letrozole include white blood cell count decreases, nausea, tiredness, diarrhea, hair thinning or hair loss, vomiting, constipation, headache, and back pain. The most common grade 3/4 side effects in the Kisqali + letrozole arm (incidence >2%) were low neutrophils, low leukocytes, abnormal liver function tests, low lymphocytes, and vomiting. Abnormalities were observed in hematology and clinical chemistry laboratory tests.

Please see the Full Prescribing Information for Kisqali, available at https://www.pharma.us.novartis.com/sites/www.pharma.us.novartis.com/files/kisqali.pdf(link is external).

About Novartis
Novartis provides innovative healthcare solutions that address the evolving needs of patients and societies. Headquartered in Basel, Switzerland, Novartis offers a diversified portfolio to best meet these needs: innovative medicines, cost-saving generic and biosimilar pharmaceuticals and eye care. Novartis has leading positions globally in each of these areas. In 2016, the Group achieved net sales of USD 48.5 billion, while R&D throughout the Group amounted to approximately USD 9.0 billion. Novartis Group companies employ approximately 118,000 full-time-equivalent associates. Novartis products are sold in approximately 155 countries around the world. For more information, please visit http://www.novartis.com.

Novartis is on Twitter. Sign up to follow @Novartis and @NovartisCancer at http://twitter.com/novartis(link is external) and http://twitter.com/novartiscancer (link is external)
For Novartis multimedia content, please visit www.novartis.com/news/media-library
For questions about the site or required registration, please contact media.relations@novartis.com

References
[1] Kisqali (ribociclib) Prescribing information. East Hanover, New Jersey, USA: Novartis Pharmaceuticals Corporation; March 2016.
[2] Novartis Data on File
[3] American Cancer Society. How Common Is Breast Cancer? Available at https://www.cancer.org/cancer/breast-cancer/about/how-common-is-breast-cancer.html(link is external). Accessed January 23, 2017.
[4] O’Shaughnessy J. Extending survival with chemotherapy in metastatic breast cancer. The Oncologist. 2005;10(Suppl 3):20-29.

рибоциклиб , ريبوسيكليب , 瑞波西利

Ribociclib « New Drug Approvals

////////////////Novartis,  Kisqali®, ribociclib, LEE011,  FDA 2017, HR+/HER2- metastatic breast cancer, рибоциклиб , ريبوسيكليب , 瑞波西利

NEW DRUG APPROVALS BLOG HITS 16 LAKH VIEWS IN 213 COUNTRIES

Trelagliptin succinate, a novel once-weekly oral dipeptidyl peptidase-4 (DPP-4) inhibitor, was approved for the Japanese market on March 26, 2015

Trelagliptin exhibited a better potency against human DPP-4 than alogliptin and sitagliptin, along with its excellent selectivity and slow-binding properties that may partially contribute to its sustained efficacy. In phase III clinical studies, once-weekly oral trelagliptin provided long-term safety and efficacy in both monotherapy and combination with other antidiabetic medicines and was proved to be noninferior to its analogue alogliptin used once daily.

2-({6-[(3R)-3-Aminopiperidin-1-yl]-3-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl}methyl)-4-fluorobenzonitrile Monosuccinate (1)

Synthesis of Trelagliptin Succinate

Shenghui Xu , Qun Hao, Hongyan Li, Zhenren Liu, and Weicheng Zhou^*

State Key Lab of New Drug & Pharmaceutical Process, Shanghai Key Lab of Anti-Infectives, Shanghai Institute of Pharmaceutical Industry, State Institute of Pharmaceutical Industry, Shanghai 201203, China

Org. Process Res. Dev., Article ASAP

DOI: 10.1021/acs.oprd.7b00013

An improved process for the synthesis of antidiabetic drug trelagliptin succinate through unprotected (R)-3-aminopiperidine was described. The impurity profile with different conditions of the key substitution was illustrated, and then the best reaction condition was identified. The optimizations also included the bromination of 4-fluoro-2-methylbenzonitrile so that the process became efficient and concise.

• 1.
  Zhang, Z.; Wallace, M. B.; Feng, J.; Stafford, J. A.; Skene, R. J.; Shi, L.; Lee, B.; Aertgeerts, K.; Jennings, A.; Xu, R.; Kassel, D. B.; Kaldor, S. W.; Navre, M.; Webb, D. R.; Gwaltney, S. L.J. Med. Chem. 2011, 54, 510– 524, DOI: 10.1021/jm101016w
• 2.
  Feng, J.; Gwaltney, S. L.; Dipeptidyl Peptidase Inhibitors. PCT Int. Appl. WO 2005095381, October 13, 2005.
• 3.

  Grimshaw, C. E.; Jennings, A.; Kamran, R.; Ueno, H.; Nishigaki, N.; Kosaka, T.; Tani, A.; Sano, H.; Kinugawa, Y.; Koumura, E.; Shi, L.; Takeuchi, K. PLoS One 2016, 11, e0157509, DOI: 10.1371/journal.pone.0157509

• 4.

  Inagaki, N.; Onouchi, H.; Maezawa, H.; Kuroda, S.; Kaku, K. Lancet Diabetes Endocrinol.2015, 3, 191– 197, DOI: 10.1016/S2213-8587(14)70251-7
• 5.

  Inagaki, N.; Sano, H.; Seki, Y.; Kuroda, S.; Kaku, K. J.Diabetes Investig. 2016, 7, 718– 726, DOI: 10.1111/jdi.12499

   6.

  Feng, J.; Gwaltney, S. L.; Dipeptidyl Peptidase Inhibitors. PCT Int. Appl. WO 2007035629, March 29, 2007.

//////////

PATENT

WO 2009074300

The synthesis starts with the formation of the 2-arylbenzimidazole derivative 6 which can be carried out starting from N-methylphenylenediamine 2 (Method A; blue path in Scheme 1) or employing o-phenylenediamine 4 in the ring closure reaction followed by N-methylation (Method B; orange path in Scheme 1). Sodium hydrogen sulfite is used to promote the condensation of the corresponding o-phenylenediamine with the Br-aromatic aldehyde 3.(6b) The next step is the coupling of the aryl bromide with isonipecotic ethyl ester in Buchwald conditions. After acidic hydrolysis with HCl under microwave irradiation, the final amide 1 was synthesized with CDI as coupling agent.

PAPER

A Scalable Route to the SMO Receptor Antagonist SEN826: Benzimidazole Synthesis via Enhanced in Situ Formation of the Bisulfite–Aldehyde Complex

A practical and scalable route to the SMO antagonist SEN826 1 is described herein, including the discussion of an alternative approach to the synthesis of the target molecule. The optimized route consists of five chemical steps. A new and efficient access to the key intermediate 6 via the bisulfite–aldehyde complex was developed, significantly enhancing the yields and reducing costs. As a result, a synthetic procedure for preparation of multihundred gram quantities of the final product has been developed.

TRIENTINE

• Molecular Formula C₆H₁₈N₄
• Average mass 146.234 Da

112-24-3 CAS

曲恩汀, KD-034, MK-0681, MK-681, TECZA, TETA, TJA-250

Aton Pharma, a subsidiary of Valeant Pharmaceuticals, has developed and launched Syprine, a capsule formulation of trientine hydrochloride, for treating Wilson disease.

Triethylenetetramine, abbreviated TETA and trien and also called trientine (INN), is an organic compound with the formula [CH₂NHCH₂CH₂NH₂]₂. This oily liquid is colorless but, like many amines, assumes a yellowish color due to impurities resulting from air-oxidation. It is soluble in polar solvents. The branched isomer tris(2-aminoethyl)amine and piperazine derivatives may also be present in commercial samples of TETA.^[1]

Trientine hydrochloride is a metal antagonist that was first launched by Merck, Sharp & Dohme in the U.S. in 1986 under the brand name Syprine for the oral treatment of Wilson’s disease.

Orphan drug designation has also been assigned in the U.S. for the treatment of patients with Wilson’s disease who are intolerant or inadequately responsive to penicillamine and in the E.U. by Univar for the treatment of Wilson’s disease

By condensation of ethylenediamine (I) with 1,2-dichloroethane (II)

Trientine hydrochloride is N,N’-bis (2-aminoethyl)-1,2-ethanediamine dihydrochloride. It is a white to pale yellow crystalline hygroscopic powder. It is freely soluble in water, soluble in methanol, slightly soluble in ethanol, and insoluble in chloroform and ether.

The empirical formula is C6H18N4·2HCI with a molecular weight of 219.2. The structural formula is:

NH₂(CH₂)₂NH(CH₂)₂NH(CH₂)₂NH₂•2HCI

Trientine hydrochloride is a chelating compound for removal of excess copper from the body. SYPRINE (Trientine Hydrochloride) is available as 250 mg capsules for oral administration. Capsules SYPRINE contain gelatin, iron oxides, stearic acid, and titanium dioxide as inactive ingredients.

Production

TETA is prepared by heating ethylenediamine or ethanolamine/ammonia mixtures over an oxide catalyst. This process gives a variety of amines, which are separated by distillation and sublimation.^[2]

Uses

The reactivity and uses of TETA are similar to those for the related polyamines ethylenediamine and diethylenetriamine. It was primarily used as a crosslinker (“hardener”) in epoxy curing.^[2]

The hydrochloride salt of TETA, referred to as trientine hydrochloride, is a chelating agent that is used to bind and remove copper in the body to treat Wilson’s disease, particularly in those who are intolerant to penicillamine. Some recommend trientine as first-line treatment, but experience with penicillamine is more extensive.^[3]

Coordination chemistry

TETA is a tetradentate ligand in coordination chemistry, where it is referred to as trien.^[4] Octahedral complexes of the type M(trien)Cl₃ can adopt several diastereomeric structures, most of which are chiral.^[5]

Trientine, chemically known as triethylenetetramine or N,N’-bis(2-aminoethyl)-l,2-ethanediamine belongs to the class of polyethylene polyamines. Trientine dihydrochloride is a chelating agent which is used to bind and remove copper in the body in the treatment of Wilson’s disease.

Trientine dihydrochloride (1)

Trientine dihydrochloride formulation, developed by Aton with the proprietary name SYPRINE, was approved by USFDA on November 8, 1985 for the treatment of patients with Wilson’s disease, who are intolerant to penicillamine. Trientine dihydrochloride, due to its activity on copper homeostasis, is being studied for various potential applications in the treatment of internal organs damage in diabetics, Alzheimer’s disease and cancer.

Various synthetic methods for preparation of triethylenetetramine (TETA) and the corresponding dihydrochloride salt have been disclosed in the prior art.

U.S. 4,806,517 discloses the synthesis of triethylenetetramine from ethylenediamine and monoethanolamine using Titania supported phosphorous catalyst while U.S. 4,550,209 and U.S. 5,225,599 disclose catalytic condensation of ethylenediamine and ethylene glycol for the synthesis of linear triethylenetetramine using catalysts like zirconium trimethylene diphosphonate, or metatungstate composites of titanium dioxide and zirconium dioxide.

U.S. 4,503,253 discloses the preparation of triethylenetetramine by reaction of an alkanolamine compound with ammonia and an alkyleneamine having two primary amino groups in the presence of a catalyst, such as supported phosphoric acid wherein the support is comprised of silica, alumina or carbon.

The methods described above for preparation of triethylenetetramine require high temperatures and pressure. Further, due to the various possible side reactions and consequent associated impurities, it is difficult to control the purity of the desired amine.

CN 102924289 discloses a process for trientine dihydrochloride comprising reduction of Ν,Ν’-dibenzyl-,N,N’-bis[2-(l,3-dioxo-2H-isoindolyl)ethyl]ethanediamine using hydrazine hydrate to give N,N’-dibenzyl-,N,N’-bis(2-aminoethyl)ethanediamine, which, upon condensation with benzyl chloroformate gave N,N’-dibenzyl-,N,N’-bis[2-(Cbz-amino)ethyl]ethanediamine, and further reductive deprotection to give the desired compound.

CS 197,093 discloses a process comprising reaction of triethylenetetramine with concentrated hydrochloric acid to obtain the crystalline tetrahydrochlonde salt. Further reaction of the salt with sodium ethoxide in solvent ethanol, filtration of the solid sodium chloride which is generated in the process, followed by slow cooling and crystallization of the filtrate provided the dihydrochloride salt. Optionally, aqueous solution of the tetrahydrochloride salt was passed through a column of an anion exchanger and the eluate containing free base was treated with a calculated amount of the tetrahydrochloride, evaporated, and the residue was crystallized from aqueous ethanol to yield the dihydrochloride salt.

The process is quite circuitous and cumbersome, requiring use of strong bases, filtration of sodium chloride and results in yields as low as 60%.

US 8,394,992 discloses a method for preparation of triethylenetetramine dihydrochloride wherein tertiary butoxycarbonyl (boc) protected triethylenetetramine is first converted to its tetrahydrochloride salt using large excess of hydrochloric acid in solvent isopropanol, followed by treatment of the resulting tetrahydrochloride salt with a strong base like sodium alkoxide to produce the amine free base (TETA) and sodium chloride salt in anhydrous conditions. The free amine is extracted with tertiary butyl methyl ether (TBME), followed by removal of sodium chloride salt and finally the amine free base TETA is treated with hydrochloric acid in solvent ethanol to give trientine hydrochloride salt.

PATENT

WO-2017046695

EXAMPLES

Example 1: Preparation of 2-([2-[cyanomethyl]-t-butyloxycarbonylamino]ethyl- 1-butyloxy carbonylamino)acetonitrile (5)

Potassium carbonate (481.9 g) was added to a stirred mixture of ethylenediamine (100.0 g) in acetonitrile (800 ml) and cooled to around 10°C. Chloroacetonitrile (263.8 g) was gradually added at same temperature and stirred at 25-30°C, till completion of the reaction, as monitored by HPLC. The mixture was cooled to 5-15°C and Boc-anhydride (762. lg) was added to it, followed by stirring at the same temperature. The temperature was raised to 25-30°C and the mass was stirred till completion of the reaction, as monitored by HPLC.

The reaction mass was filtered and the filtrate was concentrated. Toluene was added to the residue, and the mixture was heated to around 70°C followed by cooling and filtration to give 2-([2-[cyanomethyl)-t-butyloxycarbonylamino]ethyl-t-butyloxycarbonylamino) acetonitrile (5).

Yield: 506.8 g

% Yield: 89.9 %

Example 2: Preparation of t-butyl( N-2-aminoethyl)N-([2-[(2-aminoethyl)t-butyloxy)carbonylamino] ethyl) carbamate (6)

Raney nickel (120.0 g) in isopropanol (100 ml) was charged into an autoclave, followed by a mixture of Compound 5 (200 g) in isopropanol (400 ml). Cooled ammonia solution prepared by purging ammonia gas in 1400 ml isopropanol, equivalent to 125 g ammonia was gradually charged to the autoclave and the reaction was carried out around 15-25°C under hydrogen pressure of 2-5 Kg/cm².

After completion of the reaction, as monitored by HPLC, the mass was filtered, concentrated, and methyl tertiary butyl ether was added to the residue. The mixture was heated to around 50°C, followed by cooling of the mass, stirring, optional seeding with compound 6 and filtration to give tertiary butyl-(N-2-aminoethyl)N-([2-[(2-aminoethyl)-(tert-butyloxy) carbonylamino] ethyl) carbamate.

Yield: 174 g

%Yield: 85 %

Example 3: Preparation of triethylenetetramine dihydrochloride (1)

Concentrated hydrochloric acid (121.5 g) was gradually added to a stirred mixture of tertiary-butyl-N-(2-aminoethyl)-N-2-[(2-aminoethyl)-(tert-butoxy) carbonyl] amino] ethyl} carbamate (Compound 6, 200.0 g) and water (1400 ml) at 20-30°C. The reaction mixture was heated in the temperature range of 100-105°C till completion of the reaction, as monitored by HPLC, with optionally distilling out water, if so required.

The reaction mass was concentrated and ethanol (600 ml) was added to the residue, followed by heating till a clear solution was obtained. The reaction mixture was gradually cooled with stirring, filtered and dried to provide triethylenetetramine dihydrochloride (1).

Yield: 88.9 g, (70 %)

Purity : > 99%

Patent

https://www.google.com/patents/US8394992

Trientine was said to be used in the synthesis of benzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine in French Patent No. FR2810035 to Guilard et al. Cetinkaya, E., et al., “Synthesis and characterization of unusual tetraminoalkenes,” J. Chem. Soc. 5:561-7 (1992), is said to be directed to synthesis of benzylidene-(2-{3-[2-(benzylidene-amino)-ethyl]-2-phenyl-imidazolidin-1-yl}-ethyl)-amine from trientine, as is Araki T., et al., “Site-selective derivatization of oligoethyleneimines using five-membered-ring protection method,” Macromol., 21:1995-2001 (1988). Triethylenetetramine may reportedly also be used in the synthesis of N-methylated triethylenetetramine, as reported in U.S. Pat. No. 2,390,766, to Zellhoefer et al.

Synthesis of polyethylenepolyamines, including triethylenetetramines, from ethylenediamine and monoethanolamine using pelleted group IVb metal oxide-phosphate type catalysts was reported by Vanderpool et al. in U.S. Pat. No. 4,806,517. Synthesis of triethylenetetramine from ethylenediamine and ethanolamine was also proposed in U.S. Pat. No. 4,550,209, to Unvert et al. U.S. Pat. No. 5,225,599, to King et al. is said to be directed to the synthesis of linear triethylene tetramine by condensation of ethylenediamine and ethylene glycol in the presence of a catalyst. Joint production of triethylenetetramine and 1-(2-aminoethyl)-aminoethyl-piperazine was proposed by Borisenko et al. in U.S.S.R. Patent No. SU1541204. U.S. Pat. No. 4,766,247 and European Patent No. EP262562, both to Ford et al., reported the preparation of triethylenetetramine by reaction of an alkanolamine compound, an alkaline amine and optionally either a primary or secondary amine in the presence of a phosphorous containing catalyst, for example phosphoric acid on silica-alumina or Group IIIB metal acid phosphate, at a temperature from about 175° C. to 400° C. under pressure. These patents indicate that the synthetic method used therein was as set forth in U.S. Pat. No. 4,463,193, to Johnson. The Ford et al. ‘247 patent is also said to be directed to color reduction of polyamines by reaction at elevated temperature and pressure in the presence of a hydrogenation catalyst and a hydrogen atmosphere. European Patent No. EP450709 to King et al. is said to be directed to a process for the preparation of triethylenetetramine and N-(2-aminoethyl)ethanolamine by condensation of an alkylenamine and an alkylene glycol in the presence of a condensation catalyst and a catalyst promoter at a temperature in excess of 260° C.

Russian Patent No. RU2186761, to Zagidullin, proposed synthesis of diethylenetriamine by reaction of dichloroethane with ethylenediamine. Ethylenediamine has previously been said to have been used in the synthesis of N-carboxylic acid esters as reported in U.S. Pat. No. 1,527,868, to Hartmann et al.

Japanese Patent No. 06065161 to Hara et al. is said to be directed to the synthesis of polyethylenepolyamines by reacting ethylenediamine with ethanolamine in the presence of silica-treated Nb205 supported on a carrier. Japanese Patent No. JP03047154 to Watanabe et al., is said to be directed to production of noncyclic polyethylenepolyamines by reaction of ammonia with monoethanolamine and ethylenediamine. Production of non-cyclic polyethylenepolyamines by reaction of ethylenediamine and monoethanolamine in the presence of hydrogen or a phosphorous-containing substance was said to be reported in Japanese Patent No. JP03048644. Regenerative preparation of linear polyethylenepolyamines using a phosphorous-bonded catalyst was proposed in European Patent No. EP115,138, to Larkin et al.

A process for preparation of alkyleneamines in the presence of a niobium catalyst was said to be provided in European Patent No. 256,516, to Tsutsumi et al. U.S. Pat. No. 4,584,405, to Vanderpool, reported the continuous synthesis of essentially noncyclic polyethylenepolyamines by reaction of monoethanolamine with ethylenediamine in the presence of an activated carbon catalyst under a pressure between about 500 to about 3000 psig., and at a temperature of between about 200° C. to about 400° C. Templeton, et al., reported on the preparation of linear polyethylenepolyamides asserted to result from reactions employing silica-alumina catalysts in European Patent No. EP150,558.

Production of triethylenetetramine dihydrochloride was said to have been reported in Kuhr et al., Czech Patent No. 197,093, via conversion of triethylenetetramine to crystalline tetrahydrochloride and subsequently to triethylenetetramine dihydrochloride. “A study of efficient preparation of triethylenetetramine dihydrochloride for the treatment of Wilson’s disease and hygroscopicity of its capsule,” Fujito, et al., Yakuzaigaku, 50:402-8 (1990), is also said to be directed to production of triethylenetetramine.

Preparation of triethylenetetramine salts used for the treatment of Wilson’s disease was said to be reported in “Treatment of Wilson’s Disease with Triethylene Tetramine Hydrochloride (Trientine),” Dubois, et al., J. Pediatric Gastro. & Nutrition, 10:77-81 (1990); “Preparation of Triethylenetetramine Dihydrochloride for the Treatment of Wilson’s Disease,” Dixon, et al., Lancet, 1(1775):853 (1972); “Determination of Triethylenetetramine in Plasma of Patients by High-Performance Liquid Chromatography,” Miyazaki, et al., Chem. Pharm. Bull., 38(4):1035-1038 (1990); “Preparation of and Clinical Experiences with Trien for the Treatment of Wilson’s Disease in Absolute Intolerance of D-penicillamine,” Harders, et al., Proc. Roy. Soc. Med., 70:10-12 (1977); “Tetramine cupruretic agents: A comparison in dogs,” Allen, et al., Am. J. Vet. Res., 48(1):28-30 (1987); and “Potentiometric and Spectroscopic Study of the Equilibria in the Aqueous Copper(II)-3,6-Diazaoctane-1,8-diamine System,” Laurie, et al., J.C.S. Dalton, 1882 (1976).

Preparation of Triethylenetetramine Salts by Reaction of Alcohol Solutions of Amines and acids was said to be reported in Polish Patent No. 105793, to Witek. Preparation of triethylenetetramine salts was also asserted in “Polycondensation of polyethylene polyamines with aliphatic dicarboxylic acids,” Witek, et al., Polimery, 20(3):118-119 (1975).

Baganz, H., and Peissker, H., Chem. Ber., 1957; 90:2944-2949; Haydock, D. B., and Mulholland, T. P. C., J. Chem. Soc., 1971; 2389-2395; and Rehse, K., et al., Arch. Pharm., 1994; 393-398, report on Strecker syntheses. Use of Boc and other protecting groups has been described. See, for example, Spicer, J. A. et al., Bioorganic & Medicinal Chemistry, 2002; 10: 19-29; Klenke, B. and Gilbert, I. H., J. Org. Chem., 2001; 66: 2480-2483.

FIG. 6 shows an ¹H-NMR spectrum of a triethylenetetramine hydrochloride salt in D₂O, as synthesized in Example 3. NMR values include a frequency of 400.13 Mhz, a 1H nucleus, number of transients is 16, points count of 32768, pulse sequence of zg30, and sweep width of 8278.15 H

CLIP

http://jpdb.nihs.go.jp/jp17e/JP17e_1.pdf

Method of purification: Dissolve Trientine Hydrochloride in water while warming, and recrystallize by addition of ethanol (99.5). Or dissolve Trientine Hydrochloride in water while warming, allow to stand after addition of activated charcoal in a cool and dark place for one night, and filter. To the filtrate add ethanol (99.5), allow to stand in a cool and dark place, and recrystallize. Dry the crystals under reduced pressure not exceeding 0.67 kPa at 409C until ethanol odor disappears.

References

Triethylenetetramine

Names
Other names N,N’-Bis(2-aminoethyl)ethane-1,2-diamine; TETA; Trien; Trientine (INN); Syprine (brand name)
Identifiers
CAS Number	112-24-3
3D model (Jmol)	Interactive image
Beilstein Reference	605448
ChEBI	CHEBI:39501
ChemSpider	21106175
ECHA InfoCard	100.003.591
EC Number	203-950-6
Gmelin Reference	27008
KEGG	C07166
MeSH	Trientine
PubChem CID	5565
RTECS number	YE6650000
UNII	SJ76Y07H5F
UN number	2259
InChI[show]
SMILES[show]
Properties
Chemical formula	C6H18N4
Molar mass	146.24 g·mol−1
Appearance	Colorless liquid
Odor	Fishy, ammoniacal
Density	982 mg mL−1
Melting point	−34.6 °C; −30.4 °F; 238.5 K
Boiling point	266.6 °C; 511.8 °F; 539.7 K
Solubility in water	Miscible
log P	1.985
Vapor pressure	<1 Pa (at 20 °C)
Refractive index (nD)	1.496
Thermochemistry
Specific heat capacity (C)	376 J K−1 mol−1 (at 60 °C)
Pharmacology
ATC code	A16AX12 (WHO)
Hazards
GHS pictograms
GHS signal word	DANGER
GHS hazard statements	H312, H314, H317, H412
GHS precautionary statements	P273, P280, P305+351+338, P310
EU classification (DSD)	C
R-phrases	R21, R34, R43, R52/53
S-phrases	(S1/2), S26, S36/37/39, S45
Flash point	129 °C (264 °F; 402 K)
Lethal dose or concentration (LD, LC):
LD50 (median dose)	550 mg kg−1 (dermal, rabbit) 2.5 g kg−1 (oral, rat)
Related compounds
Related amines	Ethylenediamine Diethylenetriamine Cyclam
Related compounds	Ethylenediaminetetraacetic acid
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).

///////////////TRIENTINE, 112-24-3, 曲恩汀 , KD-034 , MK-0681, MK-681, TECZA, TETA, TJA-250, Orphan drug

NCCNCCNCCN

Debio-1452, AFN 1252

AFN-1252; UNII-T3O718IKKM; API-1252; CAS 620175-39-5; CHEMBL1652621; (E)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)-3-(7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl)acrylamide

GlaxoSmithKline plc INNOVATOR

Debiopharm SA,

Melioidosis, Enoyl ACP reductase Fabl inhibitor

Debio-1452, a novel class fatty acid biosynthesis (FAS) II pathway inhibitor, was studied in phase II clinical trials for the oral treatment of staphylococcal infections, including hospital and community-acquired MRSA and acute bacterial skin and skin structure infections. Debiopharm is developing oral and IV formulations of a prodrug of Debio-1452, Debio-1450.

Infections caused by or related to bacteria are a major cause of human illness worldwide. Unfortunately, the frequency of resistance to standard antibacterials has risen dramatically over the last decade, especially in relation to Staphylococcus aureus. For example, such resistant S. aureus includes MRSA, resistant to methicillin, vancomycin, linezolid and many other classes of antibiotics, or the newly discovered New Delhi metallo-beta-lactamase- 1 (NDM-1) type resistance that has shown to afford bacterial resistant to most known antibacterials, including penicillins, cephalosporins, carbapenems, quinolones and fluoroquinolones, macrolides, etc. Hence, there exists an urgent, unmet, medical need for new agents acting against bacterial targets..

In recent years, inhibitors of Fabl, a bacterial target involved in bacterial fatty acid synthesis, have been developed and many have been promising in regard to their potency and tolerability in humans, including a very promising Fabl inhibitor, (E)-N-methyl-N-((3-methylbenzofuran-2-yl)methyl)-3-(7-oxo-5,6,7,8-tetrahydro-l,8-naphthyridin-3-yl)acrylamide. This compound, however, has been found to be difficult or impracticable to formulate into acceptable oral and parenteral (e.g., intravenous or subcutaneous) formulations, and has marked insolubility, poor solution stability, and oral bioavailability. Much effort, over a decade or more, has been expended to design and synthesize an alternative compound that retains the significant inhibition of Fabl upon administration, but has improved physical and chemical characteristics that finally allow for practical oral and parenteral formulations. Up to now, no such compound has been identified that has adequate stability in the solid state, in aqueous solutions, together with excellent oral bioavailability that is necessary for oral and/or a parenteral administration, and is capable of being formulated into an oral and/or intravenous or intramuscular drug product using practical and commonly utilized methods of sterile formulation manufacture.

Debio-1452 is expected to have high potency against all drug-resistant phenotypes of staphylococci, including hospital and community-acquired MRSA.

Affinium obtained Debio-1452, also known as API-1252, through a licensing deal with GlaxoSmithKline. In 2014, Debiopharm acquired the product from Affinium.

In 2013, Qualified Infectious Disease Product designation was assigned to the compound for the treatment of acute bacterial skin and skin structure infections (ABSSSI).

SYNTHESIS

Heck coupling of 6-bromo-3,4-dihydro-1,8-naphthyridin-2-one with t-butyl acrylate in the presence of Pd(OAc)2, DIEA and P(o-tol)3  in propionitrile/DMF or acetonitrile/DMF affords naphthyridinyl-acrylate,

Whose t-butyl ester group is then cleaved using TFA in CH2Cl2 to furnish, after treatment with HCl in dioxane, 3-(7-oxo-6,8-dihydro-5H-1,8-naphthyridin-3-yl)acrylic acid hydrochloride

SEE BELOW………

Finally, coupling of acid with N-methyl-N-(3-methylbenzofuran-2-ylmethyl)amine using EDC, HOBt and DIEA in DMF provides the target AFN-1252

Preparation of N-methyl-N-(3-methylbenzofuran-2-ylmethyl)amine :

Chlorination of 3-methylbenzofuran-2-carboxylic acid  with (COCl)2 and catalytic DMF, followed by condensation with CH3NH2 in CH2Cl2 yields the corresponding benzofuran-2-carboxamide,

Which is then reduced with LiAlH4 in THF to furnish N-methyl-N-(3-methylbenzofuran-2-ylmethyl)amine.

CONTD……..

Reduction of 2-aminonicotinic acid  with LiAlH4 in THF gives (2-amino-3-pyridinyl)methanol ,

which upon bromination with Br2 in AcOH yields (2-amino-5-bromo-3-pyridinyl)methanol hydrobromide.

Substitution of alcohol  with aqueous HBr at reflux provides the corresponding bromide,

which undergoes cyclocondensation with dimethyl malonate  in the presence of NaH in DMF/THF to furnish methyl 6-bromo-2-oxo-1,2,3,4-tetrahydro-1,8-naphthyridine-3-carboxylate.

Hydrolysis of ester with NaOH in refluxing MeOH, followed by decarboxylation in refluxing HCl leads to 6-bromo-3,4-dihydro-1,8-naphthyridin-2-one

PATENT

US-20170088822

Aurigene Discovery Technologies Ltd

Novel co-crystalline polymorphic form of a binary enoyl-acyl carrier protein reductase (FabI) and FabI inhibitor ie AFN-1252. The FabI was isolated from Burkholderia pseudomallei (Bpm). The co-crystal is useful for identifying an inhibitor of FabI, which is useful for treating BpmFabI associated disease ie melioidosis. Appears to be the first patenting to be seen from Aurigene Discovery Technologies or its parent Dr Reddy’s that focuses on BpmFabI crystal; however, see WO2015071780, claiming alkylidine substituted heterocyclyl derivatives as FabI inhibitors, useful for treating bacterial infections. Aurigene was investigating FabI inhibitors, for treating infectious diseases, including bacterial infections such as MRSA infection, but its development had been presumed to have been discontinued since December 2015; however, publication of this application would suggest otherwise.

WO2015071780

PATENTS

US 20060142265

http://www.google.co.in/patents/US20060142265

PATENT

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2013190384&recNum=1&maxRec=&office=&prevFilter=&sortOption=&queryString=&tab=PCTDescription

////////////Debio-1452, AFN 1252,AFN-1252, UNII-T3O718IKKM, API-1252, 620175-39-5, PRECLINICAL, Phase 2, Qualified Infectious Disease Product designation

CC1=C(OC2=CC=CC=C12)CN(C)C(=O)C=CC3=CC4=C(NC(=O)CC4)N=C3

The greening of peptide synthesis

Abstract

The synthesis of peptides by amide bond formation between suitably protected amino acids is a fundamental part of the drug discovery process. However, the required coupling and deprotection reactions are routinely carried out in dichloromethane and DMF, both of which have serious toxicity concerns and generate waste solvent which constitutes the vast majority of the waste generated during peptide synthesis. In this work, propylene carbonate has been shown to be a green polar aprotic solvent which can be used to replace dichloromethane and DMF in both solution- and solid-phase peptide synthesis. Solution-phase chemistry was carried out with Boc/benzyl protecting groups to the tetrapeptide stage, no epimerisation occurred during these syntheses and chemical yields for both coupling and deprotection reactions in propylene carbonate were at least comparable to those obtained in conventional solvents. Solid-phase peptide synthesis was carried out using Fmoc protected amino acids on a ChemMatrix resin and was used to prepare the biologically relevant nonapeptide bradykinin with comparable purity to a sample prepared in DMF.

Boc-Ala-Phe-OBn 5a    ref S1

Boc-Ala-OH (324 mg, 1.71 mmol) and HCl.H-Phe-OBn (500 mg, 1.71 mmol) were coupled according to the general coupling procedure. The residue was purified using flash column chromatography (35:65, EtOAc:PE) to give Boc-Ala-Phe-OBn 5a as a white crystalline solid (682 mg, 93%). RF = 0.34 (40:60, EtOAc:PE);

mp 95.6-96.3 °C;

[α]D 23 -27.7 (c 1.0 in MeOH);

IR (Neat) νmax 3347 (m), 3063 (w), 3029 (w), 2928 (m), 2852 (w), 1735 (w), 1684 1666 and 1521 (s) cm-1;

1H NMR (400 MHz, CDCl3): δ = 7.36-7.31 (m, 3H, ArH), 7.29-7.24 (m, 2H, ArH), 7.26-7.21 (m, 3H, ArH), 7.04-6.97 (m, 2H, ArH), 6.72 (d J 7.7 Hz, 1H, Phe-NH), 5.16-5.10 (m, 1H, Ala-NH), 5.13 (d J 12.1 Hz, 1H, OCH2Ph), 5.07 (d J 12.1 Hz, 1H, OCH2Ph), 4.88 (dt, J 7.7, 5.9 1H, PheNCH), 4.11 (br, 1H, Ala-NCH), 3.13 (dd J 13.9, 6.1 Hz, 1H, CH2Ph), 3.08 (dd J 13.9, 6.1 Hz, 1H, CH2Ph), 1.41 (s, 9H, C(CH3)3), 1.29 (d J 6.6 Hz, 3H, CH3);

13C NMR (100 MHz, CDCl3): δ = 172.3 (C=O), 171.2 (C=O), 155.6 (NC=O), 135.7 (ArC), 135.1 (ArC), 129.5 (ArCH), 128.7 (ArCH), 128.6 (ArCH), 127.2 (ArCH), 80.2 (CMe3), 67.4 (OCH2Ph), 53.3 (Phe-NCH), 50.3 (Ala-NCH), 38.0 (CH2Ph), 28.4 (C(CH3)3), 18.5 (CH3);

MS (ESI) m/z 449 [(M+Na)+ , 100]; HRMS (ESI) m/z calculated for C24H30N2O5Na 449.2048 (M+Na)+ , found 449.2047 (0.6 ppm error).

S1 J. Nam, D. Shin, Y. Rew and D. L. Boger, J. Am. Chem. Soc., 2007, 129, 8747–8755; Q. Wang, Y. Wang and M. Kurosu, Org. Lett., 2012, 14, 3372–3375.

(1R,2S)-2-(Aminomethyl)-N,N-diethyl-1 phenylcyclobutanecarboxamide (19)

1 H NMR (CDCl3) δ 7.36–7.33 (m, 4H), 7.25–7.21 (m, 1H), 3.51–3.43 (qd, J = 13.8 Hz, 6.8 Hz, 1H), 3.15–2.87 (m, 7H), 2.81–2.72 (m, 2H), 2.23–2.14 (m, 1H), 2.04–1.97 (m, 1H), 1.62 (tdd, J = 10.5 Hz, 5.7 Hz, 2.6 Hz, 1H), 1.07 (t, J = 7.1 Hz, 3H), 0.35 (t, J = 7.1 Hz, 3H) ppm;

13C NMR (CDCl3) δ 172.7, 143.3, 128.8, 126.4, 125.3, 54.6, 44.4, 42.4, 41.0, 39.5, 31.1, 19.0, 12.2, 12.0 ppm;

IR (neat) 3364, 1622, 1437, 905, 728 cm−1 ;

[α] 20 D +1.5 (c 0.5, CHCl3) (lit.5 [α]D +0.84);

ESI-MS (ES+ ) 261 [M + H]+ ; HRMS m/z calcd for C16H25N2O: 261.1958, found: 261.1961;

chiral HPLC (CHIRALCEL OJ-RH 150 × 4.6 mm, H2O/MeOH 35 : 65, flow rate 1 mL min−1 , detection at 254 nm), tmajor = 8.5 min, tminor = 6.7 min, er 95 : 5. Of note, compound 19 was acetylated with acetic anhydride/NEt3 prior to HPLC analysis.

5 S. Cuisiat, A. Newman-Tancredi, O. Vitton and B. Vacher, WO patent, 112597, 2010

Enantioselective synthesis of a cyclobutane analogue of Milnacipran

Enantioselective synthesis of a cyclobutane analogue of Milnacipran

Dinh-Vu Nguyen,^a  Edmond Gravel,^a  David-Alexandre Buisson,^a  Marc Nicolas^b  and  Eric Doris*^a 

^aService de Chimie Bioorganique et de Marquage (SCBM), CEA, Université Paris-Saclay, 91191 Gif-sur-Yvette, France

E-mail: eric.doris@cea.fr

^bInstitut de Recherche Pierre Fabre, Centre de Recherche et Développement, 3 avenue Hubert Curien, 31035 Toulouse, France

Abstract

The asymmetric synthesis of a cyclobutane analogue of the antidepressant drug Milnacipran is reported. The optically active derivative incorporates a central cyclobutane ring in lieu of the cyclopropane unit classically found in Milnacipran. The two stereogenic centres borne by the cyclobutane were sequentially installed starting from phenylacetonitrile.

(S)-4-(2,4-Dihydroxyphenyl)-N-(1-phenylethyl)piperidine-1-carboxamide (1)

Process Development and Good Manufacturing Practice Production of a Tyrosinase Inhibitor via Titanium-Mediated Coupling between Unprotected Resorcinols and Ketones

Thibaud Gerfaud^* , Cédric Martin, Karinne Bouquet, Sandrine Talano, Corinne Millois-Barbuis, Branislav Musicki, Jean-Guy Boiteau^* , and Isabelle Cardinaud

Nestlé Skin Health R&D, 2400 Route des colles BP 87, 06902 Sophia-Antipolis Cedex, France

Org. Process Res. Dev., Article ASAP

DOI: 10.1021/acs.oprd.7b00036

*E-mail: Thibaud.Gerfaud@galderma.com., *E-mail: Jean-Guy.Boiteau@galderma.com.

ACS Editors’ Choice – This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes.

Thibaud Gerfaud

Team Leader Process Chemistry

Boiteau Jean-Guy

Head of Process Research & Development

Nestlé Skin Health

Abstract

A concise and economically attractive process for the synthesis of a novel tyrosinase inhibitor has been developed and implemented on a multikilogram scale under GMP. A major achievement to the success of the process is the development of a direct coupling between free resorcinol and ketone. First developed under basic conditions, this coupling has been turned to a novel titanium(IV) mediated process allowing good selectivity, easy isolation, and high atom efficiency. Other key steps feature an alkene reduction by palladium catalyzed transfer hydrogenation and a urea formation using N,N′-disuccinimidyl carbonate as the carbonyl source. This route allowed us to produce kilogram batches of the candidate to support preclinical and clinical studies.

Boiteau, J.-G.; Bouquet, K.; Talano, S.; Millois-Barbuis, C. Patent WO 2010/063774 A1, 2010.

More………………

Cas 1228342-28-6

MF C₂₀ H₂₄ N₂ O_3,

_{MW  340.42}

1-Piperidinecarboxamide, 4-(2,4-dihydroxyphenyl)-N-[(1S)-1-phenylethyl]-

• 4-(2,4-Dihydroxyphenyl)-N-[(1S)-1-phenylethyl]-1-piperidinecarboxamide
• 4-(2,4-Dihydroxyphenyl)piperidine-1-carboxylic acid N-((S)-1-phenylethyl)amide

Inventors	Jean-Guy Boiteau , Karine Bouquet , Sandrine Talano , Barbuis Corinne Millois
Applicant	Galderma Research & Development

WO 2010063774

Novel 4- (azacycloalkyl)benzene-l ,3-diol compounds as tyrosinase inhibitors, process for the preparation thereof and use thereof in human medicine and in cosmetics

The invention relates to novel 4- (azacycloalkyl) benzene-1, 3-diol compounds as industrial and useful products. It also relates to the process for the preparation thereof and to the use thereof, as tyrosinase inhibitors, in pharmaceutical or cosmetic compositions for use in the treatment or prevention of pigmentary disorders.

Skin pigmentation, in particular human skin pigmentation, is the result of melanin synthesis by dendritic cells, melanocytes. Melanocytes contain organelles called melanosomes which transfer melanin into the upper layers of keratinocytes which are then transported to the surface of the skin through differentiation of the epidermis (Gilchrest BA, Park HY, Eller MS, Yaar M, Mechanisms of ultraviolet light-induced pigmentation. Photochem Photobiol 1996; 63: 1-10; Hearing VJ, Tsukamoto K, Enzymatic control of pigmentation in mammals. FASEB J 1991; 5: 2902-2909) .

Among the enzymes of melanogenesis, tyrosinase is a key enzyme which catalyses the first two steps of melanin synthesis. Homozygous mutations of tyrosinase cause oculocutaneous albinism type I characterized by a complete lack of melanin synthesis (Toyofuku K, Wada I, Spritz RA, Hearing VJ, The molecular basis of oculocutaneous albinism type 1 (OCAl) : sorting failure and degradation of mutant tyrosinases results in a lack of pigmentation. Biochem J 2001; 355: 259-269) .

In order to treat pigmentation disorders resulting from an increase in melanin production, for which there is no treatment that meets all the expectations of patients and dermatologists, it is important to develop new therapeutic approaches.

Most of the skin-lightening compounds that are already known are phenols or hydroquinone derivatives.

These compounds inhibit tyrosinase, but the majority of them are cytotoxic to melanocytes owing to the formation of quinones. There is a risk of this toxic effect causing a permanent depigmentation of the skin. The obtaining of compounds that can inhibit melanogenesis while at the same time being very weakly cytotoxic or devoid of toxicity to melanocytes is most particularly sought.

Among the compounds already described in the literature, patent application WO 99/15148 discloses the use of 4-cycloalkyl resorcinols as depigmenting agents .

Patent FR2704428 discloses the use of 4-halo-resorcinols as depigmenting agents.

Patent applications WO 2006/097224 and WO 2006/097223 disclose the use of 4-cycloalkylmethyl resorcinols as depigmenting agents.

Patent application WO 2005/085169 discloses the use of alkyl 3- (2, 4-dihydroxyphenyl) propionate as a depigmenting agent.

Patent application WO 2004/017936 discloses the use of 3- (2, 4-dihydroxyphenyl) acrylamide as a depigmenting agent.

Patent application WO 2004/052330 discloses the use of 4- [ 1, 3] dithian-2-ylresorcinols as depigmenting agents .

More particularly, patent EP0341664 discloses the use of 4-alkyl resorcinols as depigmenting agents, among which 4-n-butyl resorcinol, also known as rucinol, is part of the composition of a depigmenting cream sold under the name Iklen^®.

The applicant has now discovered, unexpectedly and surprisingly, that novel compounds of 4- (azacycloalkyl) benzene-1, 3-diol structure have a very good tyrosinase enzyme-inhibiting activity and a very low cytotoxicity. Furthermore, these compounds have a tyrosinase enzyme-inhibiting activity that is greater than that of rucinol while at the same time being less cytotoxic with respect to melanocytes than rucinol.

These compounds find uses in human medicine, in particular in dermatology, and in the cosmetics field.

FR 2939135

////////////

O=C(N[C@@H](C)c1ccccc1)N2CCC(CC2)c3ccc(O)cc3O

EVP4593; EVP 4593; EVP-4593

QNZ(EVP4593) is a derivative of 6-aminoquinazoline class that has been previously isolated as an inhibitor of PMA/PHA-induced NF-κB pathway activation in Jurkat cells (IC50= 9 nM).

QNZ(EVP4593) is a derivative of 6-aminoquinazoline class that has been previously isolated as an inhibitor of PMA/PHA-induced NF-κB pathway activation in Jurkat cells (IC50= 9 nM).
IC50 Value: 9 nM [1]
Target: NF-kB signaling
in vitro: The efficacy of EVP4593 was dose-dependent in the range between 100 uM and 400 uM in the fly food. The EVP4593 had no significant effect on climbing performance of HD flies at 50 ?M. The EVP4593 had no toxic effects on Drosophila in the range of concentrations tested in our assays (50 – 400 ?M) [1]. Addition of 300 nM of EVP4593 resulted in strong attenuation of SOC Ca2+ influx in YAC128 MSN neurons. On average the amplitude of SOC Ca2+ entry in YAC128 MSN was reduced from 0.30 ± 0.02 (n = 29) in the presence of DMSO control to 0.11 ± 0.02 (n = 54) in the presence of 300 nM of EVP4593 (p < 0.001).
in vivo:

Paper

Identification of 4-N-[2-(4-phenoxyphenyl)ethyl]quinazoline-4,6-diamine as a novel, highly potent and specific inhibitor of mitochondrial complex I

Abstract

By probing the quinone substrate binding site of mitochondrial complex I with a focused set of quinazoline-based compounds, we identified substitution patterns as being critical for the observed inhibition. The structure activity relationship study also resulted in the discovery of the quinazoline 4-N-[2-(4-phenoxyphenyl)ethyl]quinazoline-4,6-diamine (EVP4593) as a highly potent inhibitor of the multisubunit membrane protein. EVP4593 specifically and effectively reduces the mitochondrial complex I-dependent respiration with no effect on the respiratory chain complexes II–IV. Similar to established Q-site inhibitors, EVP4593 elicits the release of reactive oxygen species at the flavin site of mitochondrial complex I. Recently, EVP4593 was nominated as a lead compound for the treatment of Huntingtons disease. Our results challenge the postulated primary mode-of-action of EVP4593 as an inhibitor of NF-κB pathway activation and/or store-operated calcium influx.

//////////

C1=CC=C(C=C1)OC2=CC=C(C=C2)CCNC3=NC=NC4=C3C=C(C=C4)N

GLGP 1837

CAS 1654725-02-6

MF C₁₆ H₂₀ N₄ O₃ S, MW 348.42

For cystic fibrosis treatment

1H-Pyrazole-3-carboxamide, N-[3-(aminocarbonyl)-4,7-dihydro-5,5,7,7-tetramethyl-5H-thieno[2,3-c]pyran-2-yl]-

SYNTHESIS

GLGP 1837

ABC transporters are a family of homologous membrane transporter proteins regulating the transport of a wide variety of pharmacological agents (for example drugs, xenobiotics, anions, etc…) that bind and use cellular adenosine triphosphate (ATP) for their specific activities. Some of these transporters were found to defend malignant cancer cells against chemotherapeutic agents, acting as multidrug resistance proteins (like the MDRl-P glycoprotein, or the multidrug resistance protein, MRP 1). So far, 48 ABC transporters, grouped into 7 families based on their sequence identity and function, have been identified.

ABC transporters provide protection against harmful environmental compounds by regulating a variety of important physiological roles within the body, and therefore represent important potential drug targets for the treatment of diseases associated with transporter defects, outwards cell drug transport, and other diseases in which modulation of ABC transporter activity may be beneficial.

The cAMP/ATP -mediated anion channel, CFTR, is one member of the ABC transporter family commonly associated with diseases, which is expressed in a variety of cells types, including absorptive and secretory epithelia cells, where it regulates anion flux across the membrane, as well as the activity of other ion channels and proteins. The activity of CFTR in epithelial cells is essential for the maintenance of electrolyte transport throughout the body, including respiratory and digestive tissue. (Quinton, 1990)

The gene encoding CFTR has been identified and sequenced (Kerem et al., 1989). CFTR comprises about 1480 amino acids that encode a protein made up of a tandem repeat of transmembrane domains, each containing six transmembrane helices and a nucleotide binding domain. The pair of

transmembrane domains is linked by a large, polar, regulatory (R)-domain with multiple phosphorylation sites that regulate channel activity and cellular trafficking.

Cystic fibrosis is caused by a defect in this gene which induces mutations in CFTR. Cystic fibrosis is the most common fatal genetic disease in humans, and affects -0.04% of white individuals(Bobadilla et al., 2002), for example, in the United States, about one in every 2,500 infants is affected, and up to 10 million people carry a single copy of the defective gene without apparent ill effects; moreover subjects bearing a single copy of the gene exhibit increased resistance to cholera and to dehydration resulting from diarrhea. This effect might explain the relatively high frequency of the CF gene within the population.

In contrast, individuals with two copies of the CF associated gene suffer from the debilitating and fatal effects of CF, including chronic lung infections.

In cystic fibrosis patients, mutations in endogenous respiratory epithelial CFTR fails to confer chloride and bicarbonate permeability to epithelial cells in lung and other tissues, thus leading to reduced apical anion secretion and disruptions of the ion and fluid transport. This decrease in anion transport causes an enhanced mucus and pathogenic agent accumulation in the lung triggering microbial infections that ultimately cause death in CF patients.

Beyond respiratory disease, CF patients also suffer from gastrointestinal problems and pancreatic insufficiency that result in death if left untreated. Furthermore, female subjects with cystic fibrosis suffer from decreased fertility, whilst males with are infertile.

A variety of disease causing mutations has been identified through sequence analysis of the CFTR gene of CF chromosomes (Kerem et al., 1989). AF508-CFTR, the most common CF mutation (present in at least 1 allele in~90 % of CF patients) and occurring in approximately 70% of the cases of cystic fibrosis, contains a single amino acid deletion of phenylalanine 508. This deletion prevents the nascent protein from folding correctly, which protein in turn cannot exit the endoplasmic reticulum (ER) and traffic to the plasma membrane, and then is rapidly degraded. As a result, the number of channels present in the membrane is far less than in cells expressing wild-type CFTR. In addition to impaired trafficking, the mutation results in defective channel gating. Indeed, even if AF508-CFTR is allowed to reach the cell plasma membrane by low-temperature (27°C) rescue where it can function as a cAMP-activated chloride channel, its activity is decreased significantly compared with WT-CFTR (Pasyk and Foskett, 1995).

Other mutations with lower incidence have also been identified that alter the channel regulation or the channel conductance. In case of the channel regulation mutants, the mutated protein is properly trafficked and localized to the plasma membrane but either cannot be activated or cannot function as a chloride channel (e.g. missense mutations located within the nucleotide binding domains), examples of these mutations are G551D, G178R, G1349D. Mutations affecting chloride conductance have a CFTR protein that is correctly trafficked to the cell membrane but that generates reduced chloride- flow (e.g. missense mutations located within the membrane-spanning domain), examples of these mutations are Rl 17H, R334W.

In addition to cystic fibrosis, CFTR activity modulation may be beneficial for other diseases not directly caused by mutations in CFTR, such as, for example, chronic obstructive pulmonary disease (COPD), dry eye disease, and Sjogren’s Syndrome.

[0014] COPD is characterized by a progressive and non-reversible airflow limitation, which is due to mucus hypersecretion, bronchiolitis, and emphysema. A potential treatment of mucus hypersecretion and impaired mucociliary clearance that is common in COPD could consist in using activators of mutant or wild-type CFTR. In particular, the anion secretion increase across CFTR may facilitate fluid transport into the airway surface liquid to hydrate the mucus and optimize periciliary fluid viscosity. The resulting enhanced mucociliary clearance would help in reducing the symptoms associated with COPD.

[0015] Dry eye disease is characterized by a decrease in tear production and abnormal tear film lipid, protein and mucin profiles. Many factors may cause dry eye disease, some of which include age, arthritis, Lasik eye surgery, chemical/thermal burns, medications, allergies, and diseases, such as cystic fibrosis and Sjogrens’s syndrome. Increasing anion secretion via CFTR could enhance fluid transport from the corneal endothelial cells and secretory glands surrounding the eye, and eventually improve corneal hydration, thus helping to alleviate dry eye disease associated symptoms. Sjogrens’s syndrome is an autoimmune disease where the immune system harms moisture-producing glands throughout the body, including the eye, mouth, skin, respiratory tissue, liver, vagina, and gut. The ensuing symptoms, include, dry eye, mouth, and vagina, as well as lung disease. Sjogrens’s syndrome is also associated with rheumatoid arthritis, systemic lupus, systemic sclerosis, and polymypositis/dermatomyositis. The cause of the disease is believed to lie in defective protein trafficking, for which treatment options are limited. As a consequence, modulation of CFTR activity may help hydrating the various organs and help to elevate the associated symptoms.

In addition to CF, the defective protein trafficking induced by the AF508-CFTR has been shown to be the underlying basis for a wide range of other diseases, in particular diseases where the defective functioning of the endoplasmic reticulum (ER) may either prevent the CFTR protein to exit the cell, and/or the misfolded protein is degraded (Morello et al., 2000; Shastry, 2003; Zhang et al., 2012).

[0017] A number of genetic diseases are associated with a defective ER processing equivalent to the defect observed with CFTR in CF such as glycanosis CDG type 1, hereditary emphysema (α-1-antitrypsin (PiZ variant)), congenital hyperthyroidism, osteogenesis imperfecta (Type I, II, or IV procollagen), hereditary hypofibrinogenemia (fibrinogen), ACT deficiency (α-1-antichymotrypsin), diabetes insipidus (DI), neurophyseal DI (vasopvessin hormoneN2 -receptor), neprogenic DI (aquaporin II), Charcot-Marie Tooth syndrome (peripheral myelin protein 22), Perlizaeus-Merzbacher disease, neurodegenerative diseases such as Alzheimer’s disease (APP and presenilins), Parkinson’s disease, amyotrophic lateral sclerosis, progressive supranuclear plasy, Pick’s disease, several polyglutamine neurological disorders such as Huntington’s disease, spinocerebullar ataxia type I, spinal and bulbar muscular atrophy,

dentatorubal pallidoluysian, and myotonic dystrophy, as well as spongiform encephalopathies, such as hereditary Creutzfeldt-Jakob disease (prion protein processing defect), Fabry disease (lysosomal a-galactosidase A), Straussler-Scheinker syndrome, chronic obstructive pulmonary disease (COPD), dry eye disease, and Sjogren’s Syndrome.

In addition to up-regulation of the activity of CFTR, anion secretion reduction by CFTR modulators may be beneficial for the treatment of secretory diarrheas, in which epithelial water transport is dramatically increased as a result of secretagogue activated chloride transport. The mechanism involves elevation of cAMP and stimulation of CFTR.

[0019] Regardless of the cause, excessive chloride transport is seen in all diarrheas, and results in dehydration, acidosis, impaired growth and death. Acute and chronic diarrheas remain a major medical problem worldwide, and are a significant factor in malnutrition, leading to death in children of less than five years old (5,000,000 deaths/year). Furthermore, in patients with chronic inflammatory bowel disease (IBD) and/or acquired immunodeficiency syndrome (AIDS), diarrhea is a dangerous condition

GLGP 1837

PATENT

WO 2015018823

Scheme 1: synthesis of the core and subsequent amide coupling

O

1 M HCI

amide coupling

HO Λ R-i

Example 2. Synthesis of intermediates

Intermediate 2: 2,2, 6,6-tetramethyltetrahydro-4H-pyran-4-one

Phorone or 2,6-dimethyl-2,5-heptadien-4-one (1 eq) is mixed with an aqueous 1 M HCI solution and the obtained emulsion is stirred at 40°C for 6 days. The water phase is extracted with DCM, and the organic phase is concentrated and purified by distillation to afford the desired product.

Alternative synthesis of Intermediate 2

[00208] A 20 L reactor is charged with aqueous 6M HCI and is warmed up to 30 °C. Molten Phorone is added while stirring vigorously at 40°C for up to 3 h until completion. The resulting solution is then cooled to 30°C and extracted with 4 x 1 L DCM. The combined organic phases are washed with saturated NaHC0₃ solution (400 niL) and are dried over Na₂S0₄. The resulting crude misture is then concentrated under vacuo, and finally purified by distillation.

Intermediate 3: 2-Amino-5,5, 7, 7-tetramethyl-4, 7-dihydro-5H-thieno[2, 3-c]pyran-3-carboxylic acid amide

Route 1 :

To a flask containing 2,2,6,6-tetramethyltetrahydro-4H-pyran-4-one (Int 2, 1 eq), cyanoacetamide (1 eq), sulfur (0.9 eq) and diethylamine (1.1 eq) are added. EtOH is then added and the resulting mixture is stirred at 40°C overnight. The reaction is diluted with water and partially concentrated by evaporation causing the precipitation of a solid that is separated by filtration. The cake is then washed with water and hexane to afford the desired product.

Alternative synthesis 1 of intermediate 3

Starting from 2,2,6,6-tetramethyltetrahydro-4H-pyran-4-one (Int 2, 1 eq), cyanoacetamide (1.1 eq) and morpholine (1.5 eq) are heated in EtOH at 80°C under inert atmosphere. After 6 h of heating, the mixture is cooled down, and sulfur (1.1 eq) is added. Next, the mixture is heated at 80°C overnight, then concentrated in vacuo and extracted with saturated NH₄C1 and NaHCOs. The organic phase is subsequently dried over MgSO i, filtered and concentrated in vacuo. The residue obtained can finally be purified by column chromatography.

Alternative synthesis 2 of intermediate 3

A 20L glass reactor with a mechanical stirrer (400 rpm) and a reflux condenser is charged with 2,2,6,6-tetramethyltetrahydro-4H-pyran-4-one (Int 2) (1.466 kg, 9.01 mol, l eq) and 2-cyanoacetamide (1.363 kg, 1.8 eq.) followed by absolute EtOH (4.5 L) and morpholine (0.706 kg, 0.9 eq.). The resulting suspension is heated for 23 h at 75°C (internal temperature). After 23 h, sulfur (0.26 kg, 0.9 eq.) is added in one portion at 75°C and the resulting suspension is stirred further for 90 min after which the resulting solution is cooled to 20°C. Then, the entire solution is concentrated in vacuo (50 mbar / 45°C) to yield a solid residue. Water (13.5 L) is added in one portion at 75°C and the mixture is cooled to 22°C. Stirring (700 rpm at 22°C) is continued for 2.5h. The solids are separated by filtration, dried under vacuum suction, and subsequently in the vacuum oven at 40°C over 3d to obtain yield the desired product.

Intermediate 11: Dipyrazolo l,5-a;l ‘,5’-dJpyrazine-4,9-dione

[00213] 10 g (89 mmol) of pyrrazole carboxylic acid is suspended in toluene 100 mL at room temperature. Then, 2 equivalents of thionyl chloride are added, followed by a catalytic amount of DMF (0.5 ml). The mixture was stirred for lh at 75°C. After lh at 70 °C, the reaction was cooled to room temperature, the solid material was collected by filtration, washed with toluene and resuspended in DCM. Triethylamine (2 equivalents) was added and the suspension was stirred for 2h at room temperature. The product was collected by filtration, washed with DCM and dried at 40°C under vacuum to afford the desired product.

Example 4. Illustrative examples for the Preparation of the Compounds of Invention

Compound 2: N-(3-carbamoyl-5, 5, 7, 7 -tetramet yl-5 , 7-dihydro-4H-thieno[2, 3-c]pyran-2-yl)-lH-pyr zole-5-carboxamide

[00274] Intermediate 3 (15 g, 59 mmol) and 2H-pyrazole-3-carboxylic acid (9.9 g, 88 mmol) are suspended in DCM (250 mL). Mukaiyama reagent (2-chloro-l-methylpyridinium iodide) (18.1 g, 71 mmol), TEA (24.7 mL, 177 mmol) and DMAP (3.6 g, 29 mmol) are added. The reaction mixture is stirred at 40°C overnight and then cooled. The mixture is evaporated and the obtained crude is suspended in a 1 M HC1 solution. After stirring for 10 min, the suspension is filtered and obtained precipitate is isolated. This precipitate is re-suspended in a 0.1 M citric acid solution. Again, filtration gives a precipitate. A third trituration is done using ether as a solvent to give a precipitate after filtration. Finally, the precipitate (13.6 g) is suspended in EtOH (816 mL) and heated at reflux. To this suspension, 65 mL of DMF is added and a clear solution is obtained. The solution is concentrated to 275 mL and cooled at 0°C. A suspension is obtained, the solid is separated by filtration, and the cake is dried affording the desired product.

Alternative route

[00275] To a stirred (400 rpm) solution of 600 g (2.36 mol) of Intermediate 3 in DMAc (6 L), is added at ambient temperature 1.3 equivalents of Intermediate 11. To this resulting suspension, at room temperature, DIPEA (618 mL, 1.5 eq.) is added in small portions over a period of 5 min. The resulting suspension is heated to 80 °C and stirred for 18h at this temperature. The resulting mixture is cooled to 15°C and an aqueous saturated NH₄C1 solution (7.5 L) is added over 30 minutes thus maintening the internal temperature between 15-24 °C. The resulting solid product is collected by filtration, and triturated with water (7.5 L) under mechanical stirring (600 rpm) for 30 min. The resulting suspension is filtered and the resulting solid is triturated in MTBE (8 L) under mechanical stirring for 45 minutes. The resulting solid is separated by filtration, and dried in a vacuum stove.

[00276] Finally, the solid is purified by hot trituration in ethanol. Therefore, the crude solid is suspended in absolute EtOH (16 L) for 1.5 h at 78 °C. The suspension is cooled to 20 °C and subsequently stirred for another hour. The solid product was collected by filtration, washed with 500 mL and again with 200 ml absolute EtOH, then dried to yield the desired product.

1H NMR PREDICT

13 C NMR PREDICT

REFERENCES

First speaker at 1st disclosures is Steven Van der Plas of @GalapagosNV talking about a cystic fibrosis treatment #ACSSanFran

http://acsmeetings.cenmag.org/first-time-disclosures-of-clinical-candidates-at-acssanfran/?utm_source=Facebook&utm_medium=Social&utm_campaign=MeetingSF17

//////////////GLGP 1837

NC(=O)c2c3CC(C)(C)OC(C)(C)c3sc2NC(=O)c1ccnn1