GADOBUTROL

gadolinium(III) 2,2′,2”-(10-((2R,3S)-1,3,4-trihydroxybutan-2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate

Gadobutrol, SH-L-562, Gadovist,138071-82-6

The US Food and Drug Administration (FDA) has approved Bayer HealthCare’s Gadavist (gadobutrol) injection as the first magnetic resonance contrast agent for evaluation of breast cancer in the US.

The agency has approved the new indication for Gadavist injection for intravenous use with magnetic resonance imaging of the breast to assess the presence and extent of malignant breast disease.

Approval is based on priority review of two Phase III studies with identical design (GEMMA-1 and GEMMA-2).

Bayer HealthCare’s Gadavist (gadobutrol)

Bayer’s Gadavist injection cleared for breast cancer evaluation
The US Food and Drug Administration (FDA) has approved Bayer HealthCare’s Gadavist (gadobutrol) injection as the first magnetic resonance contrast agent for evaluation of breast cancer in the US.

http://www.pharmaceutical-technology.com/news/newsbayer-gadavist-injection-cleared-breast-cancer-evaluation-4293723?WT.mc_id=DN_News

GADOBUTROL

Clinical data
AHFS/Drugs.com	International Drug Names
Licence data	US FDA:link
Pregnancy cat.	C (US)
Legal status	POM (UK) ℞-only (US)
Routes	IV
Identifiers
CAS number	138071-82-6
ATC code	V08CA09
PubChem	CID 72057
DrugBank	DB06703
UNII	1BJ477IO2L
KEGG	D07420
Chemical data
Formula	C18H31GdN4O9
Mol. mass	604.710 g/mol

………………………..

Gadobutrol (INN) (Gd-DO3A-butrol) is a gadolinium-based MRI contrast agent (GBCA).

It received marketing approval in Canada^[1] and in the United States.^[2][3][4]

As of 2007, it was the only GBCA approved at 1.0 molar concentrations.^[5]

Gadobutrol is marketed by Bayer Schering Pharma as Gadovist, and by Bayer HealthCare Pharmaceuticals as Gadavist.^[6]

WORLDCUP FOOTBALL WEEK 2014 BRAZIL

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http://www.google.com/patents/EP0988294B1?cl=en

• This type of complexes with metal ions, in particular with paramagnetic metal ions; is used for the preparation of non-ionic contrast agents for the diagnostic technique known as magnetic resonance (MRI, Magnetic Resonance Imaging), among which are ProHance^(R) (Gadoteridol, gadolinium complex of 10-(2-hydroxypropyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid), and Gadobutrol (gadolinium complex of [10-[2,3-dihydroxy-1-(hydroxymethyl)propyl]-1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid).
• [0003]
  Two different synthetic approaches are described in literature for the preparation of this kind of complexes, said approaches differing in the strategy taken to discriminate one of the four nitrogen atoms: the first one (Dischino et al., Inorg. Chem., 1991, 30, 1265 or EP 448191, EP 292689, EP 255471) is based on the selective protection of one of the nitrogen atoms by formation of the compound of formula (III), 5H,9bH-2a,4a,7-tetraazacycloocta[cd]pentalene, and on the subsequent hydrolysis to compound of formula (IV), 1-formyl-1,4,7,10-tetraazacyclododecane, followed by the carboxymethylation of the still free nitrogen atoms and by the deprotection and alkylation of the fourth nitrogen atom, according to scheme 1.
• [0004]
  The step from 1,4,7,10-tetraazacyclododecane disulfate (a commercially available product) to compound (III) is effected according to the conventional method disclosed in US 4,085,106, followed by formation of the compound of formula (IV) in water-alcohol medium.
• [0005]
  This intermediate is subsequently tricarboxymethylated with tert-butyl bromoacetate (TBBA) in dimethylformamide at 2.5°C and then treated with a toluene-sodium hydroxide diphasic mixture to give the compound of formula (V), 10-formyl-1,4,7,10-tetraazacyclododecane-1,4,7-triacetic, tris(1,1-dimethylethyl) ester, which is subsequently hydrolysed to compound of formula (II) in acidic solution.
• [0006]
  In the process described in WO 93/24469 for the synthesis of Gadobutrol, at first one of the nitrogen atoms is alkylated in conditions such as to minimize the formation of polyalkylated derivatives, then the monoalkylderivative is purified and carboxymethylated, according to scheme 2.
• [0007]
  The alkylation of 1,4,7,1,0-tetraazacyclododecane with the epoxide of formula (VI), 4,4-dimethyl-3,5,8-trioxabicyclo[5.1.0]octane, is carried out in anhydrous n-BuOH under reflux and the reaction mixture is extracted with water, evaporated to dryness and the residue is subsequently diluted with water and extracted with methylene chloride.
• [0008]
  The aqueous phase containing the mono-alkylated product (65% yield in Example 7 which reports the procedure for the preparation of 5 kg of Gadobutrol) is directly carboxymethylated at 70°C with chloroacetic acid, keeping pH 9.5 by addition of NaOH. The reaction mixture is adjusted to pH 1, concentrated to dryness and dissolved in methanol to remove the undissolved salts. The filtrate is then concentrated under vacuum, dissolved in water, and loaded onto a cation exchanger in the H⁺ form to fix the product. The subsequent elution with ammonia displaces the desired product, which is concentrated to small volume and subsequently complexed with gadolinium oxide according to conventional methods, and the resulting complex is purified by means of ion exchange resins. The overall yield is 42%.
• [0009]
  Although the first of these two processes could theoretically provide a higher yield, in that all the single steps (protection, carboxymethylation and deprotection) are highly selective, the complexity of the operations required to remove salts and solvents and to purify the reaction intermediates makes such theoretical advantage ineffective: the overall yield is in fact, in the case of Gadoteridol, slightly higher than 37%.
• [0010]
  The preparation of Gadobutrol according to the alternative process (WO 93/24469) provides a markedly better yield (72%) only on laboratory scale (example 2): example 7 (represented in the above Scheme 2) actually evidences that, when scaling-up, the yield of this process also remarkably decreases (42%).
• [0011]
  In addition to the drawback of an about 40% yield, both processes of the prior art are characterized by troublesome operations, which often involve the handling of solids, the use of remarkable amounts of a number of different solvents, some of them having undesirable toxicological or anyway hazardous characteristics.
• [0012]
  Moreover, the synthesis described by Dischino makes use of reagents which are extremely toxic, such as tert-butyl bromoacetate, or harmful and dangerous from the reactivity point of view, such as dimethylformamide dimethylacetal.
• [0013]
  An alternative to the use of dimethyl formamide dimethylacetal is suggested by J. Am. Chem. Soc. 102(20), 6365-6369 (1980), which discloses the preparation of orthoamides by means of triethyl orthoformate.
• [0014]
  EP 0596 586 discloses a process for the preparation of substituted tetraazacyclododecanes, among them compounds of formula (XII), comprising:

  • formation of the tricyclo[5.5.1.0] ring;
  • alkylation with an epoxide;
  • hydrolysis of the 10-formyl substituent;
  • reaction with an acetoxy derivative bearing a leaving group at the alpha-position.
• [0015]
  Nevertheless, this method requires quite a laborious procedure in order to isolate the product of step b).
• [0016]
  It is the object of the present invention a process for the preparation of the complexes of general formula (XII)

  wherein

  R₁ and R₂

  are independently a hydrogen atom, a (C₁-C₂₀) alkyl containing 1 to 10 oxygen atoms, or a phenyl, phenyloxy group, which can be unsubstituted or substituted with a (C₁-C₅) alkyl or hydroxy, (C₁-C₅) alkoxy, carbamoyl or carboxylic groups,

  Me³⁺

  is the trivalent ion of a paramagnetic metal;

  comprising the steps represented in the following Scheme 3:

• The process of the present invention keeps the high selectivity typical of the protection/deprotection strategy described by Dischino in the above mentioned paper, while removing all its drawbacks, thus providing for the first time a reproducible industrial process for the preparation of the concerned compounds in high yields and without use of hazardous substances.
• [0019]
  The preparation of the gadolinium complex of 10-(2-hydroxypropyl)-1,4,7,10-tetraazacyclododecane-1,4,7-tri-acetic) acid (Gadoteridol), according to scheme 4, is particularly preferred:

  in which the synthetic steps a), b), c), d), e), and f) have the meanings defined above and the epoxide of formula (XI) in step d) is propylene oxide.

• [0020]
  The preparation of the gadolinium complex of [10-[2,3-dihydroxy-1-(hydroxymethyl)propyl]-1,4,7,10-tetraazacyclododecane-1,4,7-triacetic) acid (Gadobutrol), according to the scheme 5, is also preferred.

  in which the synthetic steps a), b), c), d), e), and f) have the meanings defined above and the epoxide of formula (XI) in step d) corresponds to the one of formula (VI), defined above.

• [0021]
  On the other hand, step a) of the process of the present invention involves the use of triethyl orthoformate in the presence of an acid catalyst, instead of dialkylformamide-dialkylacetal.
• [0022]
  Triethyl orthoformate can be added in amounts ranging from 105% to 200% on the stoichiometric value.
• [0023]
  The reaction temperature can range from 110 to 150°C and the reaction time from 5 to 24 h.
• [0024]
  The catalyst is a carboxylic acid having at least 3 carbon atoms, C₃-C₁₈, preferably selected from the group consisting of propionic, butyric and pivalic acids.
• [0025]
  Triethyl orthoformate is a less toxic and less expensive product than N,N-dimethylformamide-dimethylacetal and does not involve the formation of harmful, not-condensable gaseous by-products. Moreover, triethyl orthoformate is less reactive than N,N-dimethylformamide-dimethylacetal, which makes it possible to carry out the loading procedures of the reactives as well as the reaction itself in utterly safe conditions even on a large scale, allows to better monitor the progress of the reaction on the basis of such operative parameters as time and temperature, without checking the progress by gas chromatography, and makes dosing the reactive less critical, in that it can be added from the very beginning without causing the formation of undesired by-products: all that rendering the process suitable for the production of compound (III) on the industrial scale in easily reproducible conditions.
• [0026]
  The subsequent step b) involves the carboxymethylation of compound (III) in aqueous solution, using a haloacetic acid, to give compound (IX), i.e. the 10-formyl-1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid salt with an alkali or alkaline-earth metal, the salts of compound (IX) with sodium, potassium or calcium being most preferred.

Example 2

• [0065]
• [0066]
  The procedure of Example 1 is followed until step C included, to obtain a solution of DO3A trisodium salt.
• [0067]
  pH is adjusted to 12.3 with conc. HCl and 57.7 kg (0.4 kmol) of 4,4-dimethyl-3,5,8-trioxabicyclo[5.1.0]-octane are added. After reaction for 4 h at 40°C and for 8 h at 80°C, the solution is cooled to 50°C, 120 kg of an aqueous solution containing 0.135 kmol of gadolinium trichloride are added. After 1 h the mixture is cooled at 17°C and acidified to pH 1.7 with conc. HCl, keeping this pH for 2 h. The solution is subsequently warmed to 50°C, pH is adjusted to 7 with sodium hydroxide, keeping these conditions for 1 h.
• [0068]
  After that, the resulting crude Gadobutrol is purified repeating exactly the same process as in steps E and F of Example 1.

Recovery of the product (Gadobutrol)

• [0069]
  The product-rich fraction is then thermally concentrated to a viscous residue and the residue is added with 350 kg of ethanol at 79°C.
• [0070]
  The resulting suspension is refluxed for 1 h, then cooled, centrifuged and dried under reduced pressure to obtain 66.0 kg of Gadobutrol (0.109 kmol), HPLC assay 99.5% (A%).
  Overall yield: 79.1%
• [0071]
  The IR and MS spectra are consistent with the indicated structure.

References

Graphical abstract: Alternative solid-state forms of a potent antimalarial aminopyridine: X-ray crystallographic, thermal and solubility aspects

Alternative solid-state forms of a potent antimalarial aminopyridine: X-ray crystallographic, thermal and solubility aspects

Dyanne L. Cruickshank, Yassir Younis, Nicholas M. Njuguna, Dennis S. B. Ongarora, Kelly Chibale and Mino R. Caira

CrystEngComm, 2014, 16, 5781 DOI:10.1039/C3CE41798K

http://pubs.rsc.org/en/Content/ArticleLanding/2014/CE/C3CE41798K?utm_medium=email&utm_campaign=pub-CE-vol-16-issue-26&utm_source=toc-alert#!divAbstract

3-(6-Methoxypyridin-3-yl)-5-(4-methylsulfonyl phenyl)-pyridin-2-amine (MMP) is a member of a novel class of orally active antimalarial drugs. This aminopyridine molecule has shown potent in vitro antiplasmodial activity and in vivo antimalarial activity in Plasmodium berghei-infected mice. The aqueous solubility of this molecule is, however, limited.

Thus investigations aimed at improving the physicochemical properties, including solubility, of MMP were accordingly conducted. Five salts of MMP were formed with co-former molecules saccharin, salicylic acid, fumaric acid, oxalic acid and suberic acid, but a cocrystal was obtained when the co-former adipic acid was employed.

All these new multi-component systems have been fully characterised using X-ray diffraction and thermal methods. Semi-quantitative, turbidimetric solubility tests in a phosphate-buffered saline solution at a pH of 7.4 were performed on the salts and the cocrystal of MMP. The saccharinate salt, fumarate salt and the cocrystal of MMP proved to have greater solubility than MMP itself. This work illustrates the importance of screening and modifying candidate drug compounds in their preliminary stages of development.
Alternative solid-state forms of a potent antimalarial aminopyridine: X-ray crystallographic, thermal and solubility aspects

Dyanne L. Cruickshank,a Yassir Younis,a Nicholas M. Njuguna,a Dennis S. B. Ongarora,a Kelly Chibalea and Mino R. Caira*a
*corresponding authors
aDepartment of Chemistry, University of Cape Town, Rondebosch 7701, South Africa
E-mail: mino.caira@uct.ac.za;
Fax: +27 21 650 5195 ;
Tel: +27 21 650 3071

CrystEngComm, 2014,16, 5781-5792

DOI: 10.1039/C3CE41798K

Haloprogin

Haloprogin is a topical antifungal agent. Its structure does not contain any of the functional groups typically exploited in hydrogen bond based co-crystal design. On the other hand, its 1-iodoalkyne moiety is nicely tailored to a crystal engineering strategy based on halogen bonding. Here we describe the formation of three polymorphs of haloprogin and of three co-crystals that this active pharmaceutical ingredient forms with both neutral and ionic co-crystal formers. The halogen bond plays a major role in all of the six structures and the interaction is thus confirmed to be a valuable tool which may complement the hydrogen bond when polymorphs and co-crystals of active pharmaceutical ingredients are pursued.

Graphical abstract: Polymorphs and co-crystals of haloprogin: an antifungal agent

http://pubs.rsc.org/en/Content/ArticleLanding/2014/CE/C4CE00367E?utm_medium=email&utm_campaign=pub-CE-vol-16-issue-26&utm_source=toc-alert#!divAbstract
Polymorphs and co-crystals of haloprogin: an antifungal agent

Michele Baldrighi,a Davide Bartesaghi,a Gabriella Cavallo,a Michele R. Chierotti,b Roberto Gobetto,b Pierangelo Metrangolo,*a Tullio Pilati,a Giuseppe Resnati*a and Giancarlo Terraneo*a
*Corresponding authors
aNFMLab-Laboratory of Nanostructured Fluorinated Materials (NFMLab), Department of Chemistry, Materials, and Chemical Engineering “Giulio Natta”, Politecnico di Milano, Via L. Mancinelli 7, 20131 Milan, Italy
E-mail: pierangelo.metrangolo@polimi.it, giuseppe.resnati@polimi.it, giancarlo.terraneo@polimi.it
bDepartment of Chemistry, Università di Torino, Via P. Giuria 7, 10125 Torino, Italy

CrystEngComm, 2014,16, 5897-5904

DOI: 10.1039/C4CE00367E

EXTRA INFO

Systematic (IUPAC) name
1,2,4-trichloro-5-[(3-iodoprop-2-yn-1-yl)oxy]benzene
Clinical data
Legal status	Not available in U.S.
Routes	Topical
Identifiers
CAS number	777-11-7
ATC code	D01AE11
PubChem	CID 3561
DrugBank	DB00793
ChemSpider	3440
UNII	AIU7053OWL
KEGG	D00339
ChEMBL	CHEMBL1289
Chemical data
Formula	C9H4Cl3IO
Mol. mass	361.39 g/mol
Physical data
Melt. point	113.5 °C (236 °F)
Solubility in water	insoluble mg/mL (20 °C)

Haloprogin is an antifungal drug used to treat athlete’s foot and other fungal infections. It is marketed in creams under the trade names Halotex, Mycanden, Mycilan, and Polik.

Action

Haloprogin was previously used in 1% topical creams as an antifungal agent. It was marketed over the counter primarily to treat tinea infections of the skin. The mechanism of action is unknown.^[1]

Haloprogin had a high incidence of side effects including: irritation, burning, vesiculation (blisters), scaling, and itching. It has since been discontinued due to the emergence of more modern antifungals with fewer side effects.^[2]

Haloprogin is used as a topical ointment or cream in the treatment of Tinea infections. Tinea infections are superficial fungal infections caused by three species of fungi collectively known as dermatophytes (Trichophyton, Microsporum and Epidermophyton). Commonly these infections are named for the body part affected, including tinea corporis (general skin), tinea cruris (groin), and tinea pedis (feet). Haloprogin is a halogenated phenolic ether administered topically for dermotaphytic infections. The mechanism of action is unknown, but it is thought to be via inhibition of oxygen uptake and disruption of yeast membrane structure and function. Haloprogin is no longer available in the United States and has been discontinued.

U.S. Patent 3,322,813.

References

1. “Haloprogin”. Drugs@FDA. Food and Drug Administration. Retrieved 2007-02-17.
2. “Haloprogin”. DrugBank. University of Alberta. Nov 6, 2006. Retrieved 2007-02-17.

A unique public-private collaboration among the National Cancer Institute (NCI), part of the National Institutes of Health, SWOG Cancer Research, Friends of Cancer Research, the Foundation for the National Institutes of Health (FNIH), five pharmaceutical companies (Amgen, Genentech, Pfizer, AstraZeneca, and AstraZeneca’s global biologics R&D arm, MedImmune), and Foundation Medicine today announced the initiation of the Lung Cancer Master Protocol (Lung-MAP) trial.

http://www.cancer.gov/newscenter/newsfromnci/2014/LungMAPlaunch?utm_content=sf27390795&utm_medium=spredfast&utm_source=facebook&utm_campaign=National+Cancer+Institute&cid=sf27390795

Rose Bengal disodium

4 ,5,6,7-Tetrachloro-2′,4′,5′,7′-tetraiodo-3-oxo-3H-spiro[2-benzofuran-1,9'-xanthene]-3′,6′-diolate disodium salt

C20 H2 Cl4 I4 O5 . 2 Na,  mw 1017.36, PH 10

http://www.talkmarkets.com/content/stocks–equities/provectus-phase-iii-melanoma-trial-results-earlier-than-planned?post=44803

The FDA has granted PV-10 Orphan Drug Status for the treatment of highly lethal metastatic melanoma and metastatic liver cancer. It has a successful and expanding Compassionate Use Program in operation and successfully completed trials on metastatic cancer of the breast, liver and melanoma, with positive results in all three. Positive effects in this context is that, if you inject PV-10 into a solid tumor, it kills cancer cells, usually within a week and doesn’t harm normal tissue. Many injected tumors actually disappear while others shrink and stop growing. The dual action of the drug is that the destruction of the cancer by direct injection of PV-10 serves to sensitize the patient’s immune system to seek out and kill similar cancer throughout the body. There is convincing evidence that untreated cancer distant from the treated cancer is attacked by the patient’s immune system after treatment.

PROVECTUS COMPANY OVERVIEW

Provectus (PVCT) is a clinical stage bio-pharmaceutical drug development company. There are 3 key scientific managers running the business along with the CFO, who is also the Chief Operating Officer. They preside over a stable of expert and specialized consultants. The company has two lead drug candidates: PH-10 for significant, often severe, and common skin disorders and PV-10, a dual action, local ablation and immunological anti-cancer drug. PH-10 is currently the subject of post-Phase II trial research into mode of action. PV-10 has successfully completed Phase II trials for malignant melanoma, is currently the subject of independent research on mode of actioRose Bengal disodium is in early clinical trials at Provectus for the topical treatment of psoriasis and atopic dermatitis. An intralesional injectable formulation is also in early clinical development as breast cancer, liver cancer and melanoma therapy. Development for the treatment of actinic keratosis had been ongoing; however, no recent development for this indication has been reported. The company is seeking approval in the U.S. to begin clinical evaluation of this formulation for the treatment of liver and prostate cancer. A compassionate use program is under way for Rose Bengal disodium for the treatment of non-visceral cancers.

The drug’s mechanism of action is believed to be characterized by the creation of free radicals upon activation, which eliminate diseased cells. The compound concentrates in tumors at cytotoxic levels while quickly dissipating from healthy tissue. Simultaneously, the drug triggers an immune response that can eliminate metastatic tumor tissue.

In 2007, orphan drug designation was assigned to Rose Bengal disodium by the FDA for the treatment of metastatic melanoma. This designation was also assigned to the compound in the U.S. in 2011 for the treatment of hepatocellular carcinoma.n and efficacy in conjunction with radiation, and it will have a Phase III pivotal trial starting shortly.

SUMMARY

1. If PV-10 and the Chemotherapies act as the prior data indicate, an NDA for melanoma may be submitted by Provectus in the first half of 2015.

2. If this occurs, the FDA denial of the Breakthrough Therapy designation will not have slowed PV-10′s progress to commercialization.

3. Given the relative safety and efficacy of the different drugs, if the trial is not stopped very early for humanitarian reasons, the planned Interim Analysis is likely to result in the cancellation of the trial, prior to the end of 2015.

4. Given PV-10′s superior safety and lack of significant side effects, if it is only as good as Chemotherapy, it will deserve FDA approval.

The Phase III pivotal trial will demonstrate the safety and efficacy of PV-10 to the market and to prospective acquirers a lot earlier than many have presumed.

Rose bengal (4,5,6,7-tetrachloro-2′,4′,5′,7′-tetraiodofluorescein) is a stain. Its sodium salt is commonly used in eye drops to stain damaged conjunctival and corneal cells and thereby identify damage to the eye. The stain is also used in the preparation of Foraminifera for microscopic analysis, allowing the distinction between forms that were alive or dead at the time of collection.

A form of Rose Bengal is also being studied as a treatment for certain cancers and skin conditions. The cancer formulation of the drug, known as PV-10, is currently undergoing clinical trials for melanoma and breast cancer. The company also has formulated a drug based on Rose Bengal for the treatment of eczema and psoriasis; this drug, PH-10, is currently in clinical trials as well.

Rose bengal
IUPAC name[hide] 4,5,6,7-Tetrachloro-3′,6′-dihydroxy-2′,4′,5′,7′-tetraiodo-3H- spiro[isobenzofuran-1,9'-xanthen]-3-one
Other names C.I. 45440
Identifiers
CAS number	11121-48-5
ATC code	S01JA02
Jmol-3D images	Image 1
SMILES [show]
Properties
Molecular formula	C20H4Cl4I4O5
Molar mass	973.67 g mol−1

Chemical applications

Light microscopy image of the undescribed species of Spinoloricus from Loricifera stained with Rose Bengal.

Rose Bengal is also used in synthetic chemistry to generate singlet oxygen from triplet oxygen. The singlet oxygen can then undergo a variety of useful reactions, particularly [2 + 2] cycloadditions with alkenes and similar systems.

Rose Bengal can be used to form many derivatives that have important medical functions. One such derivative was created so to be sonosensative but photoinsensative, so that with a high intensity focused ultrasound, it could be used in the treatment of cancer. The derivative was formed by amidation of Rose Bengal, which turned off the fluorescent and photosensitive properties of Rose Bengal, leading to a usable compound, named in the study as RB2.^[1]

Salts of Rose Bengal can also be formed, with the molecular formula C20 H4 Cl4 I4 O5 . 2 Na, molecular weight of 1017.64 g/mol and CAS # 632-69-9. Known as Rose Bengal Sodium Salt, this compound has its own unique uses and properties, but also functions as a dye.^[2]

Biological applications

PV-10 was found to cause an observable response in 60 percent of tumors treated, according to researchers in a phase II melanoma study. Locoregional disease control was observed in 75 percent of patients. Also confirmed was a “bystander effect”, previously observed in the phase I trial, whereby untreated lesions responded to treatment as well, potentially due to immune system response. These data were based on the interim results of the first 40 patients treated in an 80 patient study.^[3] Rose Bengal has been shown to not just prevent the growth and spread of ovarian cancer, but also to cause apoptotic cell death of the cancer cells. This has been proven in vitro, in order to prove that Rose Bengal is still a possible option in the treatment of cancer, and further research should be done.^[4]

Rose Bengal is also used in animal models of ischemic stroke (photothrombotic stroke models) in biomedical research. A bolus of the compound is injected into the venous system. Then the region of interest (e.g., the cerebral cortex) is exposed and illuminated by LASER light of 561 nm. A thrombus is formed in the illuminated blood vessels, causing a stroke in the dependent brain tissue.^[5][6]

Rose bengal has been used for 50 years to diagnose liver and eye cancer. It has also been used as an insecticide.^[7][8]

Rose Bengal is able to stain cells whenever the surface epithelium is not being properly protected by the preocular tear film, because Rose Bengal has been proven to not be able to stain cells because of the protective functioning of these preocular tear films.^[9] This is why Rose Bengal is often useful as a stain in diagnosing certain medical issues, such as conjunctival and lid disorders.^[10]

Rose Bengal has been used for ocular surface staining to study the efficacy of punctal plugs in the treatment of keratoconjunctivitis sicca. ^[11]

Rose Bengal is being researched as an agent in creating nano sutures.^[12] Wounds are painted on both sides with it and then illuminated with an intense light. This links the tiny collagen fibers together sealing the wound.^[13][14][15] Healing is faster and the seal reduces chances of infection.^[16][17]

Rose Bengal is used in several microbiological media, including Cooke’s Rose Bengal agar, to suppress bacterial growth.

Rose Bengal has been used as a protoplasm stain to discriminate between living and dead micro-organisms, particularly Foraminifera, since the 1950s when Bill Walton developed the technique.^[18]

Electronic applications

Rose Bengal demonstrates at least six distinct electronic properties^[19] which are otherwise hidden in the molecule. Rose Bengal is a double planar molecule and relative rotation of the planes generate unique electronics. Therefore, Rose Bengal is a suitable candidate for molecular electronics.

History

Rose Bengal was originally prepared in 1884 by Gnehm, as an analogue of fluorescein.^[20] The name is due to its similarity to alta, a dye that women in Bengal have used for centuries to colour their feet red during weddings and festivals.

References

External links

• Rose Bengal at the US National Library of Medicine Medical Subject Headings (MeSH)
• Absorption and extinction data

http://www.orphan-drugs.org/2014/06/17/selexipag-meets-primary-endpoint-pivitol-phase-iii-griphon-outcome-study-patients-pulmonary-arterial-hypertension/#sthash.PI3PzVZd.dpbs

http://www1.actelion.com/sites/en/scientists/development-pipeline/phase-3/selexipag.page

ABOUT THE ACTELION / NIPPON SHINYAKU ALLIANCE

Actelion and Nippon Shinyaku entered into an exclusive worldwide alliance in April 2008 to collaborate on selexipag, a first-in-class orally-available, selective IP receptor agonist for patients suffering from pulmonary arterial hypertension (PAH). This compound was originally discovered and synthesized by Nippon Shinyaku. Phase II evaluation has been completed, and a Phase III program in PAH patients has been initiated. Actelion is responsible for global development and commercialization of selexipag outside Japan, while the two companies will co-develop and co-commercialize in Japan. Nippon Shinyaku will receive milestone payments based on development stage and sales milestones as well as royalties on any sales of selexipag.

Selexipag
IUPAC name 2-{4-[(5,6-diphenylpyrazin-2-yl)(propan-2-yl)amino]butoxy}-N-(methanesulfonyl)acetamide
Identifiers
CAS number	475086-01-2
PubChem	9913767
ChemSpider	8089417
UNII	5EXC0E384L
KEGG	D09994
Jmol-3D images	Image 1
SMILES [show]
InChI [show]
Properties
Molecular formula	C26H32N4O4S
Molar mass	496.6 g·mol−1

NS-304 (ACT-293987), an orally available long acting non-prostanoid prostaglandin I2 (PGI-2) receptor agonist, is in phase III clinical trials at Actelion for the oral treatment of pulmonary hypertension. Nippon Shinyaku is conducting phase III clinical trials with NS-304 for this indication in Europe. In Japan, phase II clinical trials are ongoing for the treatment of pulmonary hypertension and chronic thromboembolic pulmonary hypertension.

Originally discovered and synthesized by Nippon Shinyaku, NS-304 stimulates PGI-2 receptors in blood vessels and exerts vasodilating effects.

In 2008, the compound was licensed to Actelion by Nippon Shinyaku on a worldwide basis with the exception of Japan for the oral treatment of pulmonary arterial hypertension (PAH). According to the final licensing agreement, Actelion will be responsible for global development and commercialization of NS-304 outside Japan, while the two companies will codevelop and co-commercialize the product candidate in Japan. In 2005, orphan drug designation was assigned in the E.U. by Nippon Shinyaku for the treatment of pulmonary arterial hypertension and chronic thromboembolic pulmonary hypertension.

…………………….

US2012/101276

http://www.google.st/patents/US20120101276?hl=pt-PT&cl=en

The present invention relates to a crystal of 2-{4-[N-(5,6-diphenylpyrazin-2-yl)-N-isopropylamino]butyloxy}-N-(methylsulfonyl)acetamide (hereinafter referred to as “compound A”).

BACKGROUND OF THE INVENTION

Compound A has an excellent PGI2 agonistic effect and shows a platelet aggregation inhibitory effect, a vasodilative effect, a bronchodilative effect, a lipid deposition inhibitory effect, a leukocyte activation inhibitory effect, etc. (see, for example, in WO 2002/088084 (“WO ’084”)).

Specifically, compound A is useful as preventive or therapeutic agents for transient ischemic attack (TIA), diabetic neuropathy, diabetic gangrene, peripheral circulatory disturbance (e.g., chronic arterial occlusion, intermittent claudication, peripheral embolism, vibration syndrome, Raynaud’s disease), connective tissue disease (e.g., systemic lupus erythematosus, scleroderma, mixed connective tissue disease, vasculitic syndrome), reocclusion/restenosis after percutaneous transluminal coronary angioplasty (PTCA), arteriosclerosis, thrombosis (e.g., acute-phase cerebral thrombosis, pulmonary embolism), hypertension, pulmonary hypertension, ischemic disorder (e.g., cerebral infarction, myocardial infarction), angina (e.g., stable angina, unstable angina), glomerulonephritis, diabetic nephropathy, chronic renal failure, allergy, bronchial asthma, ulcer, pressure ulcer (bedsore), restenosis after coronary intervention such as atherectomy and stent implantation, thrombocytopenia by dialysis, the diseases in which fibrosis of organs or tissues is involved [e.g., Renal diseases (e.g., tuburointerstitial nephritis), respiratory diseases (e.g., interstitial pneumonia (pulmonary fibrosis), chronic obstructive pulmonary disease), digestive diseases (e.g., hepatocirrhosis, viral hepatitis, chronic pancreatitis and scirrhous stomachic cancer), cardiovascular diseases (e.g, myocardial fibrosis), bone and articular diseases (e.g, bone marrow fibrosis and rheumatoid arthritis), skin diseases (e.g, cicatrix after operation, scalded cicatrix, keloid, and hypertrophic cicatrix), obstetric diseases (e.g., hysteromyoma), urinary diseases (e.g., prostatic hypertrophy), other diseases (e.g., Alzheimer's disease, sclerosing peritonitis; type I diabetes and organ adhesion after operation)], erectile dysfunction (e.g., diabetic erectile dysfunction, psychogenic erectile dysfunction, psychotic erectile dysfunction, erectile dysfunction associated with chronic renal failure, erectile dysfunction after intrapelvic operation for removing prostata, and vascular erectile dysfunction associated with aging and arteriosclerosis), inflammatory bowel disease (e.g., ulcerative colitis, Crohn’s disease, intestinal tuberculosis, ischemic colitis and intestinal ulcer associated with Behcet disease), gastritis, gastric ulcer, ischemic ophthalmopathy (e.g., retinal artery occlusion, retinal vein occlusion, ischemic optic neuropathy), sudden hearing loss, avascular necrosis of bone, intestinal damage caused by administration of a non-steroidal anti-inflammatory agent (e.g., diclofenac, meloxicam, oxaprozin, nabumetone, indomethacin, ibuprofen, ketoprofen, naproxen, celecoxib) (there is no particular limitation for the intestinal damage so far as it is damage appearing in duodenum, small intestine and large intestine and examples thereof include mucosal damage such as erosion and ulcer generated in duodenum, small intestine and large intestine), and symptoms associated with lumbar spinal canal stenosis (e.g., paralysis, dullness in sensory perception, pain, numbness, lowering in walking ability, etc. associated with cervical spinal canal stenosis, thoracic spinal canal stenosis, lumbar spinal canal stenosis, diffuse spinal canal stenosis or sacral stenosis) etc. (see, for example, in WO ’084, WO 2009/157396, WO 2009/107736, WO 2009/154246, WO 2009/157397, and WO 2009/157398).

In addition, compound A is useful as an accelerating agent for angiogenic therapy such as gene therapy or autologous bone marrow transplantation, an accelerating agent for angiogenesis in restoration of peripheral artery or angiogenic therapy, etc. (see, for example, in WO ’084).

Production of Compound A

Compound A can be produced, for example, according to the method described in WO ’084, and, it can also be produced according to the production method mentioned below.

Step 1:

6-Iodo-2,3-diphenylpyrazine can be produced from 6-chloro-2,3-diphenylpyrazine by reacting it with sodium iodide. The reaction is carried out in the presence of an acid in an organic solvent (e.g., ethyl acetate, acetonitrile, acetone, methyl ethyl ketone, or their mixed solvent). The acid to be used is, for example, acetic acid, sulfuric acid, or their mixed acid. The amount of sodium iodide to be used is generally within a range of from 1 to 10 molar ratio relative to 6-chloro-2,3-diphenylpyrazine, preferably within a range of from 2 to 3 molar ratio. The reaction temperature varies depending on the kinds of the solvent and the acid to be used, but may be generally within a range of from 60° C. to 90° C. The reaction time varies depending on the kinds of the solvent and the acid to be used and on the reaction temperature, but may be generally within a range of from 9 hours to 15 hours.

Step 2:

5,6-Diphenyl-2-[(4-hydroxybutyl(isopropyl)amino]pyrazine can be produced from 6-iodo-2,3-diphenylpyrazine by reacting it with 4-hydroxybutyl(isopropyl)amine. The reaction is carried out in the presence of a base in an organic solvent (e.g., sulfolane, N-methylpyrrolidone, N,N-dimethylimidazolidinone, dimethyl sulfoxide or their mixed solvent). The base to be used is, for example, sodium hydrogencarbonate, potassium hydrogencarbonate, potassium carbonate, sodium carbonate or their mixed base. The amount of 4-hydroxybutyl(isopropyl)amine to be used may be generally within a range of from 1.5 to 5.0 molar ratio relative to 6-iodo-2,3-diphenylpyrazine, preferably within a range of from 2 to 3 molar ratio. The reaction temperature varies depending on the kinds of the solvent and the base to be used, but may be generally within a range of from 170° C. to 200° C. The reaction time varies depending on the kinds of the solvent and the base to be used and on the reaction temperature, but may be generally within a range of from 5 hours to 9 hours.

Step 3:

Compound A can be produced from 5,6-diphenyl-2-[4-hydroxybutyl(isopropyl)amino]pyrazine by reacting it with N-(2-chloroacetyl)methanesulfonamide. The reaction is carried out in the presence of a base in a solvent (N-methylpyrrolidone, 2-methyl-2-propanol or their mixed solvent). The base to be used is, for example, potassium t-butoxide, sodium t-butoxide or their mixed base. The amount of N-(2-chloroacetyl)methanesulfonamide to be used may be generally within a range of from 2 to 4 molar ratio relative to 5,6-diphenyl-2-[4-hydroxybutyl(isopropyl)amino]pyrazine, preferably within a range of from 2 to 3 molar ratio. The reaction temperature varies depending on the kinds of the solvent and the base to be used, but may be generally within a range of from −20° C. to 20° C. The reaction time varies depending on the kinds of the solvent and the base to be used and on the reaction temperature, but may be generally within a range of from 0.5 hours to 2 hours.

The compounds to be used as the starting materials in the above-mentioned production method for compound A are known compounds, or can be produced by known methods.

…………………………………

WO 2002088084

and

http://www.google.fm/patents/WO2009157398A1?cl=en

………………………

Bioorganic and Medicinal Chemistry, 2007 ,  vol. 15,   21  p. 6692 – 6704

compd 31

……………………

Bioorganic and Medicinal Chemistry, 2007 ,  vol. 15,   24  p. 7720 – 7725

2a isthe drug

N-Acylsulfonamide and N-acylsulfonylurea derivatives of the carboxylic acid prostacyclin receptor agonist 1 were synthesized and their potential as prodrug forms of the carboxylic acid was evaluated in vitro and in vivo. These compounds were converted to the active compound 1 by hepatic microsomes from rats, dogs, monkeys, and humans, and some of the compounds were shown to yield sustained plasma concentrations of 1 when they were orally administered to monkeys. These types of analogues, including NS-304 (2a), are potentially useful prodrugs of 1.

http://www.sciencedirect.com/science/article/pii/S0968089607007614

References

1. Kuwano et al. NS-304, an orally available and long-acting prostacyclin receptor agonist prodrug. J Pharmacol Exp Ther 2007;322:1181-1188.
2. Kuwano et al. A long-acting and highly selective prostacyclin receptor agonist prodrug, NS-304, ameliorates rat pulmonary hypertension with unique relaxant responses of its active form MRE-269 on rat pulmonary artery. J Pharmacol Exp Ther 2008;326:691-699.
3. Simonneau G, Lang I, Torbicki A, Hoeper MM, Delcroix M, Karlocai K, Galie N. Selexipag, an oral, selective IP receptor agonist for the treatment of pulmonary arterial hypertension Eur Respir J 2012; 40: 874-880
4. Mubarak KK. A review of prostaglandin analogs in the management of patients with pulmonary arterial hypertension. Respir Med 2010;104:9-21.
5. Sitbon, O.; Morrell, N. (2012). “Pathways in pulmonary arterial hypertension: The future is here”. European Respiratory Review 21 (126): 321–327. doi:10.1183/09059180.00004812. PMID 23204120.

Technetium (^99mTc) tilmanocept……………….CAS1262984-82-6

[99mTc]-DTPA-mannosyl-dextran
Systematic (IUPAC) name
Dextran 3-[(2-aminoethyl)thio]propyl 17-carboxy-10,13,16-tris(carboxymethyl)-8-oxo-4-thia-7,10,13,16-tetraazaheptadec-1-yl 3-[[2-[[1-imino-2-(D-mannopyranosylthio)ethyl]amino]ethyl]thio]propyl ether technetium-99m complexes…………………………………………………..………………..OTHER NAME ………………Dextran 3-[(2-aminoethyl)thio]propyl 17-carboxy-10,13,16-tris(carboxymethyl)-8-oxo-4-thia-7,10,13,16-tetraazaheptadec-1-yl 3-[[2-[[1-imino-2-(D-mannopyranosylthio)ethyl]amino]ethyl]thio]propyl ether technetium-99Tc complexes (1-6)-alpha-D-pyranoglucan partially etherified by 3-[(2-aminoethyl)sulfanyl]propyl 17-carboxy-10,13,16-tris(carboxymethyl)-8-oxo-4-thia-7,10,13,16-tetraazaheptadecyl and 3-[(2-{[2-(L-mannopyranosylsulfanyl)acetimidoyl]amino}ethyl)sulfanyl]propyl [99mTc]technetium coordination compound [99mTc]-DTPA-mannosyl-dextran composed of a dextran backbone linked to multiple units of mannose and DTPA (diethylenetriamine pentaacetic acid) with an average molecular weight of 35800………………..LAUNCHED………….Launched – 2013
Clinical data
Trade names	Lymphoseek
AHFS/Drugs.com	entry
Pregnancy cat.	C (US)
Legal status	℞-only (US)
Routes	Intradermal, subcutaneous
Pharmacokinetic data
Half-life	1.75 to 3.05 hours at injection site
Identifiers
ATC code	V09IA09
Chemical data
Formula	(C6H10O5)n(C19H28N4O9S99mTc)3–8(C13H24N2O5S2)12–20(C5H11NS)0–17
	Mol. mass 15,281–23,454 g/mol[1]……………………..CODES1600 NEO3-06 TcDTPAmanDx Tilmanocepthttp://chem.sis.nlm.nih.gov/chemidplus/rn/1262984-82-6NDA N202207 APPROVED Mar 13, 2013

PATENT US 6409990, EXPMay 12, 2020

商品名：Lymphoseek  通用名：Technetium Tc 99m tilmanocept  中文名：未知
药企：Navidea Biopharmaceuticals, Inc.

FDA approves Navidea’s Lymphoseek for expanded use in head and neck cancer patients

The US Food and Drug Administration (FDA) has approved Navidea Biopharmaceuticals’ Supplemental New Drug Application (sNDA) for the expanded use of Lymphoseek (technetium Tc 99m tilmanocept) Injection indicated for guiding sentinel lymph node (SLN) biopsy in head and neck cancer patients with squamous cell carcinoma of the oral cavity.

http://drugdelivery.pharmaceutical-business-review.com/news/fda-approves-navideas-lymphoseek-for-expanded-use-in-head-and-neck-cancer-patients-160614-4294154

NCI: 99mTc-DTPA-mannosyl-dextran A radiolabeled macromolecule consisting of the chelating agent diethylenetriamine pentaacetic acid (DTPA) and mannose each attached to a dextran backbone and labeled with metastable technetiumTc-99 (Tc-99m), with mannose binding and radioisotopic activities. Upon injection, the mannose moiety of 99mTc-DTPA-mannosyl-dextran binds to mannose-binding protein (MBP). As MBPs reside on the surface of dendritic cells and macrophages, this gamma-emitting macromolecule tends to accumulate in lymphatic tissue where it may be imaged using gamma scintigraphy. This agent exhibits rapid clearance from the injection site, rapid uptake and high retention within the first draining lymph node, and low uptake by the remaining lymph nodes. MBP is a C-type lectin that binds mannose or fucose carbohydrate residues, such as those found on the surfaces of many pathiogens, and once bound activates the complement system.

The active ingredient in technetium Tc 99m tilmanocept is technetium Tc 99m tilmanocept. The active ingredient is formed when Technetium Tc 99m pertechnetate, sodium injection is added to the tilmanocept powder vial.

Technetium Tc 99m binds to the diethylenetriaminepentaacetic acid (DTPA) moieties of the tilmanocept molecule.

Chemically, technetium Tc 99m tilmanocept consists of technetium Tc 99m, dextran 3-[(2- aminoethyl)thio]propyl 17-carboxy-10,13,16- tris(carboxymethyl)-8-oxo-4-thia-7,10,13,16- tetraazaheptadec-1-yl 3-[[2-[[1-imino-2-(D- mannopyranosylthio) ethyl]amino]ethyl]thio]propyl ether complexes. Technetium Tc 99m tilmanocept has the following structural formula:

Empirical formula: [C₆H₁₀O₅]_n.(C₁₉H₂₈N₄O₉S^99mTc)_b.(C₁₃H₂₄N₂O₅S₂)_c.(C₅H₁₁NS)_a

Calculated average molecular weight: 15,281 to 23,454 g/mol

It contains 3-8 conjugated DTPA (diethylenetriamine pentaacetic acid) molecules (b); 12-20 conjugated mannose molecules (c) with 0-17 amine side chains (a) remaining free.

The tilmanocept powder vial contains a sterile, non-pyrogenic, white to off-white powder that consists of a mixture of 250 mcg tilmanocept, 20 mg trehalose dihydrate, 0.5 mg glycine, 0.5 mg sodium ascorbate, and 0.075 mg stannous chloride dihydrate. The contents of the vial are lyophilized and are under nitrogen.

Technetium Tc 99m tilmanocept injection is supplied as a Kit. The Kit includes tilmanocept powder vials which contain the necessary non-radioactive ingredients needed to produce technetium Tc 99m tilmanocept. The Kit also contains DILUENT for technetium Tc 99m tilmanocept. The diluent contains a preservative and is specifically formulated for technetium Tc 99m tilmanocept. No other diluent should be used.

The DILUENT for technetium Tc 99m tilmanocept contains 4.5 mL sterile buffered saline consisting of 0.04% (w/v) potassium phosphate, 0.11% (w/v) sodium phosphate (heptahydrate), 0.5% (w/v) sodium chloride, and 0.4% (w/v) phenol. The pH is 6.8 – 7.2.http://www.druginformation.com/RxDrugs/T/Technetium%20Tc%2099m%20Tilmanocept%20Injection.html

Lymphoseek(TM) is a lymphatic tissue-targeting agent which was first launched in 2013 in the U.S. by Navidea Biopharmaceuticals (formerly known as Neoprobe) for lymphatic mapping with a hand-held gamma counter to assist in the localization of lymph nodes draining a primary tumor site in patients with breast cancer or melanoma. In 2014, a supplemental NDA was approved in the U.S. for its use as a sentinel lymph node tracing agent in patients with head and neck squamous cell carcinoma of the oral cavity. Although several tracing agents exist that are used in “off-label” capacities, Lymphoseek is the first tracing agent specifically labeled for lymph node detection.

In 2012, an MAA was filed in the E.U. for the detection of lymphatic tissue in patients with solid tumors, and in 2013, a supplemental MAA was filed in the E.U. for sentinel lymph node detection in patients with head and neck cancer. The products is also awaiting registration to support broader and more flexible use in imaging and lymphatic mapping procedures, including lymphoscintigraphy and other optimization capabilities.

Navidea holds an exclusive worldwide license of Lymphoseek(TM) through the University of California at San Diego (UCSD), and, in 2007, Lymphoseek(TM) was licensed to Cardinal Health by Navidea for marketing and distribution in the U.S.

Lymphoseek(TM), also known as [99mTc]DTPA-mannosyl-dextran, is a receptor-binding radiopharmaceutical designed specifically for the mapping of sentinel lymph nodes in connection with gamma detection devices in a surgical procedure known as intraoperative lymphatic mapping (ILM). It is made up of multiple DTPA and mannose units, each attached by a 5-carbon thioether spacer to a dextran backbone. The compound features subnanomolar affinity for the mannose binding protein receptor, and consequently shows low distal node accumulation. Additionally, its small molecular diameter of 7 nanometers allows for enhanced diffusion into lymphatic channels and capillaries.

1600
99mTc-tilmanocept
Tc-DTPA-mannosyl-dextran
Technetium Tc 99m Tilmanocept
Tilmanocept
UNII-8IHI69PQTC

Chemical structure of [99mTc]tilmanocept. [99mTc]Tilmanocept is composed of a dextran backbone (black) to which are attached multiple units of mannose (green) and DTPA (blue). The mannose units provide a molecular mechanism by which [99mTc]tilmanocept avidly binds to a receptor specific to reticuloendothelial cells (CD206), and the DTPA units provide a highly stable means to radiolabel tilmanocept with 99mtechnetium (red). The molecular weight of [99mTc]tilmanocept is approximately 19,000 g/mol; the molecular diameter is 7.1 nm

[(99m)Tc]Tilmanocept is a CD206 receptor-targeted radiopharmaceutical designed for sentinel lymph node (SLN) identification. Two nearly identical nonrandomized phase III trials compared [(99m)Tc]tilmanocept to vital blue dye.

Technetium (^99mTc) tilmanocept, trade name Lymphoseek, is a radiopharmaceutical diagnostic imaging agent approved by the U.S. Food and Drug Administration (FDA) for the imaging of lymph nodes.^[1][2] It is used to locate those lymph nodes which may be draining from tumors, and assist doctors in locating those lymph nodes for removal during surgery.^[3]

http://blog.sina.com.cn/u/1242475203

…………………….

WO 2000069473

http://www.google.com/patents/EP1178838A2?cl=en

…………………………………………..

US 6409990

 http://www.google.co.in/patents/US6409990

References

1. FDA Professional Drug Information
2. http://www.fda.gov/NewsEvents/Newsroom/PressAnnouncements/ucm343525.htm
3. Marcinow, A. M.; Hall, N.; Byrum, E.; Teknos, T. N.; Old, M. O.; Agrawal, A. (2013). “Use of a novel receptor-targeted (CD206) radiotracer, 99mTc-tilmanocept, and SPECT/CT for sentinel lymph node detection in oral cavity squamous cell carcinoma: Initial institutional report in an ongoing phase 3 study”. JAMA otolaryngology– head & neck surgery 139 (9): 895–902. doi:10.1001/jamaoto.2013.4239. PMID 24051744.

http://www.google.com/patents/US8247538

Radiopharmaceuticals for use in therapy employ radionuclides which are generally longer in half-life and weaker in penetration capability, but emit stronger radiation, sufficient to kill cells, in relation to that for use in diagnosis. Alpha ray-emitting radionuclides are excluded from radiopharmaceuticals for the reason that they are highly radioactive and difficult to purchase and to attach to other compounds. All of the radionuclides currently used in pharmaceuticals are species that emit beta rays.

As mentioned above, radiopharmaceuticals, whether for use in therapy or diagnosis, are prepared by labeling pharmaceuticals with specific radionuclides. Technetium-99m (^99mTc) is known as the radioisotope most widely used to label radiopharmaceuticals. Technetium-99m has a half life of as short as 6 hours and emits gamma rays at 140 KeV, and thus it is not so toxic to the body. In addition, gamma radiation from the radioisotope is highly penetrative enough to obtain images. Thanks to these advantages, technetium-99m finds a broad spectrum of therapeutic and diagnostic applications in the nuclear medicine field (Sivia, S. J., John, D. L., Potential technetium small molecule radiopharmaceuticals. Chem. Rev. 99, 2205-2218, 1999; Shuang, L., Edwards, D. S., ^99mTc-Labeled small peptides as diagnostic radiopharmaceuticals. Chem. Rev. 99, 2235-2268, 1999).

Methods of labeling ^99mTc-2,6-diisopropylacetanilidoiminodiacetic acid are well known in the art (Callery, P. S., Faith, W. C., et al., 1976. Tissue distribution of technetium-99m and carbon-labeled N-(2,6)-dimetylphenylcarbamoylmethyl iminodiacetic acid. J. Med. Chem. 19, 962-964; Motter, M. and Kloss, G., 1981. Properties of various IDA derivatives. J. Label. Compounds Padiopharm. 18, 56-58; Cao, Y. and Suresh, M. R. 1998. A Simple And Efficient Method For Radiolabeling Of Preformed Liposomes. J Pharm Pharmaceut Sci. 1 (1), 31-37).

Basically, the conventional methods are based on the following reaction formula. In practice, a solution of SnCl₂.2H₂O, serving as a reducing agent of technetium-99m, in 0.1 N HCl and 0.1 ml (10 mCi) of sodium pertechnetium were added to lyophilized 2,6-diisopropylacetanilidoiminodiacetic acid in a vial, followed by stirring at room temperature for 30 min to prepare ^99mTc-2,6-diisopropylacetanilidoiminodiacetic acid. The preparation of ^99mTc-2,6-diisopropylacetanilidoiminodiacetic acid may be realized according to the following reaction formula.

 

 

Such conventional processes of preparing radiopharmaceuticals labeled with technetium-99m can be divided into reactions between the radioisotope and a physiologically active material to be labeled and the separation of labeled compounds from unlabeled compounds.

	M. Molter, et al., Properties of Various IDA Derivatives, J. Label. Compounds Padiopharm., vol. 18, pp. 56-58, 1981.
2		Patrick S. Callery, et al., Tissue Distribution of Technetium-99m and Carbon . . . , J. Med. Chem., vol. 19, pp. 962-964, 1976.
3	*	Sang Hyun Park et al. Synthesis and Radiochemical Labeling of N-(2,6-diisopropylacetanilido)-Iminodiacetic acid and it s analogues under microwave irradiation: A hepatobiliary imaging agent, QSAR Comb. Sci. 2004, 23, 868-874.
4		Shuang Liu, et al., 99mTc-Labeled Small Peptides as Diagnostic . . . , Chem. Rev., vol. 99, pp. 2235-2268, 1999.
5		Shuang Liu, et al., 99mTc—Labeled Small Peptides as Diagnostic . . . , Chem. Rev., vol. 99, pp. 2235-2268, 1999.
6		Silvia S. Jurisson, et al., Potential Technetium Small Molecule . . . , Chem. Rev., vol. 99, pp. 2205-2218, 1999.
7		Y. Cao, et al., A Simple and Efficient Method for Radiolabeling . . . , J. Phar,. Pharmaceut. Sci., pp. 31-37, 1998.

 

 

 

Diagnostic radiopharmaceuticals (V09)
Central nervous system	99mTc (Exametazime) 123I (Ioflupane Iofetamine Iomazenil) 18F (Florbetapir)
Skeletal system	99mTc (Medronic acid)
Renal	123I (Sodium iodohippurate)
Hepatic/reticuloendothelial	75Se (SeHCAT)
Respiratory system	133Xe 81mKr
Cardiovascular system	99mTc (Sestamibi Tetrofosmin) 111In (Imciromab) 82Rb (Rubidium chloride)
Inflammation/infection	99mTc (Exametazime Sulesomab Tilmanocept) 111In 67Ga
Tumor	99mTc (Arcitumomab Votumumab Hynic-octreotide) 111In (Capromab pendetide Satumomab pendetide) 125I (CC49) 123I/131I (Iobenguane) 18F (Fludeoxyglucose Sodium fluoride)
Adrenal cortex	123I/125I (Iodocholesterol)
Radionuclides (including tracers)	positron (PET): 18F 11C 13N 15O 68Ga gamma ray/photon (SPECT/scintigraphy): 99mTc 123I 125I 131I 111In 57Co 153Sm 133Xe 51Cr 81mKr 201Tl 67Ga 75Se

N’-(5,6,7,8-Tetrahydroquinolin-8-ylidene)-4-(2-pyridyl)piperazine-1-carbothiohydrazide

http://criticaloutcome.com/110819_COTI-2%20Fact%20Sheet.pdf

http://www.slideshare.net/trevorheisler/about-coti2

MW 366.483, C19 H22 N6 S

Caspase 9 Activators
PKB beta/Akt2 Inhibitors

Critical Outcome Technologies,

Critical Outcome Technologies (COTI) (Originator)  preclinical for ovary cancer

http://drugdiscovery.pharmaceutical-business-review.com/news/critical-outcome-technologies-receives-orphan-drug-designation-for-coti-2-180614-4296271

Critical Outcome Technologies has announced that the US Food and Drug Administration (FDA) has granted COTI-2 an Orphan Drug Designation for the treatment of ovarian cancer.
Critical Outcome Technologies president and CEO Dr Wayne Danter said that receiving the Orphan Drug Designation for COTI-2 speaks to the need for new treatment options for patients with ovarian cancer.

http://drugdiscovery.pharmaceutical-business-review.com/news/critical-outcome-technologies-receives-orphan-drug-designation-for-coti-2-180614-4296271

• COTI-2 | A Potential Breakthrough Therapy for Many Cancers June 11, 2013
• About COTI-2 Late preclinical drug candidate discovered using CHEMSAS® – the company’s proprietary, artificial intelligence-based drug discovery technology 2
• COTI-2 highlights 1 Potential breakthrough therapy for many cancers 2 Active against many cancers with mutations of the p53 gene 3 > 50% of all human cancers have a p53 mutation 3
• Why p53 is important? p53 is a tumour suppressing gene If mutated, cancers can develop & grow without control A mutation of the p53 gene is the most common mutation found in human cancer cells 4
• The future of cancer treatments COTI-2 targets and primarily destroys tumor cells Traditional chemotherapy kills growing & dividing cells, cancer or healthy COTI-2 would treat genetic mutations common in many types of cancer Most current treatments are organ specific (i.e. treatment for lung cancer, colon cancer, etc.) 5
• COTI-2 development progress Easily synthesized oral formulation with no stability issues Effective alone or in combination with approved cancer drugs In final two-species toxicity studies prior to FDA filing enabling human trials 6

http://www.google.com/patents/WO2008083491A1?cl=en

EXAMPLES

Synthesis of COTI-2 The synthesis of COTI-2, as depicted above, was conducted according to the following synthetic methodology:

DCM R T

H₂N-NH₂

lmidazol-1 -yl-(4-pyridin-2-yl-piperazin-1 -yl)-methanethione (or intermediate 3 above) was formed as follows. Λ/-(2-pyridyl) piperazine (MW 163.22, 0.91 ml, 6.0 mmoles, 1 eq) 2 was added to a solution of 1 ,1 ‘- thiocarbonyldiimidazole (MW 178.22, 1.069 g, 6.0 mmoles, 1 eq) 1 in 50 ml of dichloromethane at room temperature. The reaction mixture was stirred overnight at room temperature. The mixture was washed with water, dried \ over sodium sulfate, filtered and concentrated to provide imidazol-1-yl-(4- pyridin-2-yl-piperazin-1-yl)-methanethione (MW 273.36, 1.354 g, 4.95 mmol, 83% yield) 3, which was used without further purification. TLC (CH₂CI₂/MeOH: 95/5): Rf = 0.60, Product UV and Ninhydrin stain active. ¹H-NMR (400 MHz, CDCI₃), δ ppm: 3.72 (s, 4H), 4.02 (s, 4H), 6.67 (d, 1 H, J = 7 Hz), 6.72 (dd, 1 H, J = 7 and 5 Hz), 7.11 (s, 1 H), 7.24 (s, 1 H), 7.54 (t, 1 H, J = 7 Hz), 7.91 (s, 1 H), 8.20 (d, 1 H, J = 5 Hz).

Hydrazine hydrate (MW 50.06, 0.26 ml, 5.44 mmoles, 1.1 eq) was added to a solution of imidazol-1-yl-(4-pyridin-2-yl-piperazin-1-yl)- methanethione 3 (MW 210.30, 1.040 g, 4.95 mmol, 1 eq) in 30 ml of ethanol at room temperature. The reaction mixture was stirred under reflux for 2 hours. A white precipitate formed. This white solid was filtered off and rinsed with diethyl ether to yield 1-[Λ/-(2-pyridyl)-piperazine)-carbothioic acid hydrazide (MW 237.33, 0.86 g, 3.62 mmol, 73% yield) 4 as a white solid, and used without further purification. TLC (CH₂CI₂/MeOH: 95/5): Rf = 0.20, Product UV and Ninhydrin stain active. ¹H-NMR (400 MHz, DMSO-d₆), δ ppm: 3.53 (s, 4H), 3.85 (s, 4H), 6.66 (dd, 1 H, J = 8 and 5 Hz), 6.82 (d, 1 H, J = 8 Hz), 7.55 (t, 1 H, J = 8 Hz), 8.12 (d, 1 H, J = 5 Hz).

COTI-2

Finally, COTI-2 was formed as follows. 1-[Λ/-(2-pyridyl)-piperazine)- carbothioic acid hydrazide (MW 237.33, 0.475 g, 2.0 mmol, 1 eq) 4 and 6,7- dihydro-5H-quinolin-8-one (MW 147.18, 0.306 g, 2.0 mmol, 1 eq) 5 was dissolved in 15 ml of ethanol at room temperature. The mixture was then stirred under reflux for 20 hours. A yellow solid precipitated out of the solution. This solid was filtered off then rinsed with methanol and diethyl ether to yield COTI-2 (MW 366.48, 0.60 g, 1.64 mmol, 82% yield) as a yellow solid. TLC (CH₂CI₂/MeOH: 95/5): Rf = 0.75, Product UV and Ninhydrine stain active. HPLC analysis showed a mixture of isomers (approximately in 80/20 ratio), and >98% purity. During the HPLC Method Development, as expected, this product tends to be hydrolyzed in presence of TFA in mobile phase solution. MS (ESI+, 0.025% TFA in 50/50 MeOH/H₂O): [M+H]⁺ = 367.1 , [M+Na]⁺ = 389.1 ; ¹H-NMR (400 MHz, CDCI₃), δ ppm (Major isomer): 2.09 (m, 2H), 2.92 (m, 4H), 3.67 (m, 4H), 4.27 (m, 4H), 6.69 (dd, 1 H, J = 8 and 5 Hz)₁ 7.25 (dd,

1 H, J = 8 and 5 Hz), 7.55 (d, 2H, J = 8 Hz), 8.23 (d, 1 H, J = 5 Hz), 8.63 (d, 1 H, \ J = 5 Hz), 14.76 (s, 1 H). δ ppm (Minor isomer): 2.09 (m, 2H), 3.14 (t, 4H, J = 6 Hz), 3.80 (m, 4H), 4.27 (m, 4H), 6.66 (m, 1 H), 7.31 (dd, 1 H, J = 8 and 5 Hz), 7.52 (m, 1 H), 7.70 (d, 1 H, J = 8 Hz), 8.23 (d, 1 H, J = 5 Hz), 8.53 (d, 1 H, J = 5 Hz), 15.65 (s, 1 H).

…………………………..

BMS-791325, Beclabuvir

IN PHASE 2 for Hepatitis C (HCV)

An NS5B inhibitor.

BMS-791325 preferably is

CAS

958002-33-0
958002-36-3 (as hydrochloride)

C36 H45 N5 O5 S, 659.838

Cycloprop(d)indolo(2,1-a)(2)benzazepine-9-carboxamide, 12-cyclohexyl-N-((dimethylamino)sulfonyl)-4b,5,5a,6-tetrahydro-3-methoxy-5a-((3-methyl-3,8-diazabicyclo(3.2.1)oct-8-yl)carbonyl)-, (4bS,5aR)-

(4bS,5aR)-12-Cyclohexyl-N-(dimethylsulfamoyl)-3-methoxy-5a-((3-methyl-3,8-diazabicyclo(3.2.1)oct-8-yl)carbonyl)-4b,5,5a,6-tetrahydrocyclopropa(d)indolo(2,1-a)(2)benzazepine-9-carboxamide

(4bS,5aR)-12-Cyclohexyl-N-(dimethylsulfamoyl)-3-methoxy-5a-((3-methyl-3,8-diazabicyclo(3.2.1)oct-8-yl)carbonyl)-4b,5,5a,6-tetrahydrocyclopropa(d)indolo(2,1-a)(2)benzazepine-9-carboxamide

(1aR,12bS)-8-Cyclohexyl-N-(dimethylsulfamoyl)-11-methoxy-1a-[(3-methyl-3,8-diazabicyclo[3.2.1]oct-8-yl)carbonyl]-1,1a,2,12b-tetrahydrocyclopropa[d]indolo[2,1-a][2]benzazepine-5-carboxamide

Cycloprop [d] indolo [2, 1 -a] [2]benzazepine-5-carboxamide, 8-cyclohexyl-N- [(dimethylamino)sulfonyl]-l,la,2,12b-tetrahydro-ll-methoxy-la-[(3-methyl-3,8- diazabicyclo[3.2.1]oct-8-yl)carbonyl]-, (laR,12bS)-

Bristol-Myers Squibb (Originator)

RNA-Directed RNA Polymerase (NS5B) Inhibitors

UNII-MYW1X5CO9S

BMS-791325 is in phase II clinical studies at Bristol-Myers Squibb for the treatment of chronic hepatitis C. In 2013, the company received breakthrough therapy designation in the U.S. for the treatment of chronic hepatitis C in combination with daclatasvir and asunaprevir.

Squibb Bristol Myers Co,

http://www.google.com/patents/WO2007136982A1?cl=en

Scheme 1.

N-protected piperazines can also be coupled to the intermediate indolobenzazepine acids and the resultant piperazine carboxamides can be deprotected using methods known in the art and derivatized using a variety of synthetic protocols, some illustrative examples of which are shown below (See Scheme 2).

Scheme 2.

An intermediate useful for the synthesis of some compounds of the invention involves the preparation of the tert-butyl ester indolobenzazepine shown in Scheme 3. Scheme 3.

t-Butylation either:

This methodology involves base catalyzed hydrolysis of the indole methyl ester shown, followed by its reaction with either thionyl chloride and potassium tertiary butoxide, or alkylation with silver carbonate and tertiary butyl bromides. The resultant compound can be transformed using chemistry analogous to that outlined previously to provide the mixed ester indolobenzazepines shown above.

Scheme 4.

Some examples exist as stereoisomeric mixtures. The invention encompasses all stereoisomers of the compounds. Methods of fractionating stereoisomeric mixtures are well known in the art, and include but are not limited to; preparative chiral supercritical fluid chromatography (SFC) and chiral high performance liquid chromatography (HPLC). An example using this approach is shown in scheme 5. Scheme 5.

An additional method to achieve such separations involves the preparation of mixtures of diastereomers which can be separated using a variety of methods known in the art. One example of this approach is shown below (Scheme 6).

Scheme 6.

Diastereomers separated by reverse phase HPLC

Some diastereomeric amides can be separated using reverse phase HPLC. After hydroysis, the resultant optically active acids can be coupled with bridged piperazine derivatives (Scheme 6). For example, O-(lH-benzotriazol-l-yl)-N,N, N’,N’-tetramethyluronium tetrafluoroborate and diisopropyl ethyl amine in DMSO can be used to give the alkyl bridged piperazine carboxamides. Other standard acid amine coupling methods can also be used to give optically active carboxamides.

Schemes 7-9 illustrate other methods of making intermediates and compounds.

Scheme 8.

Scheme 9.

Biological Methods

The compounds demonstrated activity against HCV NS5B as determined in the following HCV RdRp assays.

DESCRIPTION OF SPECIFIC EMBODIMENTS

Unless otherwise specified, analytical LCMS data on the following intermediates and examples were acquired using the following columns and conditions. Stop time: Gradient time + 1 minute; Starting cone: 0% B unless otherwise noted; Eluent A: 5% CH₃CN / 95% H₂O with 10 mM NH₄OAc (for columns A, D and E); 10 % MeOH / 90 % H₂O with 0.1% TFA (for columns B and C); Eluent B: 95% CH₃CN / 5% H₂O with 10 mM NH₄OAc (for columns A, D and E); 90 % MeOH / 10 % H₂O with 0.1% TFA (for columns B and C); Column A:

Phenomenex lOμ 4.6 x 50 mm C18; Column B: Phenomenex C18 lOμ 3.0 x 50 mm; Column C: Phenomenex 4.6 x 50 mm C18 lOμ; Column D: Phenomenex Lina C18 5μ 3.0 x 50 mm; Column E: Phenomenex 5μ 4.6 x 50 mm Cl 8.

Intermediate 1

lH-Indole-6-carboxylic acid, 2-bromo-3-cyclohexyl-, methyl ester. Freshly recrystallized pyridinium tribromide (recrystallization from hot AcOH (5 mL per 1 g), rinsed with cold AcOH and dried under high vacuum over KOH) was added in portions (over 10 min.) to a stirring solution of methyl 3-cyclohexyl-lH-indole-6- carboxylate (60 g, 233 mmol) (prepared using procedures describe in WO2004/065367) in CHC1/THF (1: 1, 1.25 L) at 2o C. The reaction solution was stirred at 0-5 °C for 2.5h, and washed with sat. aq. NaHSO₃ (1 L), 1 N HCl (1 L) and brine (1 L). The organic layer was dried (MgSO₄) and concentrated. The resulting red oil was diluted with Et₂θ and concentrated. The resulting pink solid was dissolved into Et2θ (200 mL) treated with hexanes (300 mL) and partially concentrated. The solids were collected by filtration and rinsed with hexanes. The mother liquor was concentrated to dryness and the procedure repeated. The solids were combined to yield lH-indole-6-carboxylic acid, 2-bromo-3-cyclohexyl-, methyl ester (64 g, 190 mmol, 82%) as a fluffy pink solid, which was used without further purification. IHNMR (300 MHz, CDCl₃) δ 8.47 (br s, IH), 8.03 (d, J = 1.4 Hz, IH), 7.74 (dd, J = 1.4, 8.8 Hz, IH), 7.69 (d, J = 8.8 Hz, IH), 3.92 (s, 3H), 2.82 (tt, J = 3.7, 11.7 Hz, IH), 1.98 – 1.72 (m, 7H), 1.50 – 1.27 (m, 3H). 13CNMR (75 MHz, CDC13) δ 168.2, 135.6, 130.2, 123.1, 120.8, 120.3, 118.7, 112.8, 110.7, 52.1, 37.0, 32.2(2), 27.0(2), 26.1. LCMS: m/e 334 (M-H)^“, ret time 3.34 min, column A, 4 minute gradient.

Intermediate 2

lH-Indole-6-carboxylic acid, 2-bromo-3-cyclohexyl-. A solution of methyl 2- bromo-S-cyclohexyl-lH-indole-ό-carboxylate (20 g, 60 mmol) and LiOH (3.8 g, 160 mmol) in MeOΗ/TΗF/Η₂O ( 1 : 1 : 1 , 300 mL) was heated at 90 °C for 2h. The reaction mixture was cooled in an ice/H₂O bath, neutralized with IM HCl (-160 mL) diluted with H₂O (250 mL) and stirred for Ih at rt. The precipitates were collected by filtration rinse with H₂O and dried to yield lH-indole-6-carboxylic acid, 2-bromo-3- cyclohexyl- (quant.) which was used without further purification.

An alternative procedure that can by used to provide lH-indole-6-carboxylic acid, 2-bromo-3-cyclohexyl- is described below: A solution of methyl 2-bromo-3-cyclohexyl-lH-indole-6-carboxylate (117 g, 349 mmol) and LiOKH₂O (26.4 g, 629 mmol) in MeOH/THF/H2O (1: 1: 1, 1.8 L) was heated at reflux for 3h. The reaction mixture was cooled in an ice/H2O bath to ~2 °C, neutralized with IM HCl (-650 mL) (added at such a rate that temperature did not exceed 5 °C), diluted with H2O (1 L) and stirred while warming to ambient temperature. The precipitates were collected by filtration rinsed with H₂O and dried to yield the mono THF solvate of lH-indole-6-carboxylic acid, 2-bromo-3- cyclohexyl- (135.5 g, 345 mmol, 99%) as a yellow solid, which was used without further purification. IHNMR (300 MHz, CDCl₃) δ 11.01 (br s, IH), 8.77 (s, IH), 8.07 (d, J = 1.5 Hz, IH), 7.82 (dd, J = 1.5, 8.8 Hz, IH), 7.72 (d, J = 8.8 Hz, IH), 3.84 – 3.74 (m, 4H), 2.89 (m, IH), 1.98 – 1.72 (m, HH), 1.50 – 1.24 (m, 3H). 13CNMR (75 MHz, CDC13) δ 172.7, 135.5, 130.7, 122.3, 120.9(2), 118.8, 113.3, 111.1, 67.9(2), 37.0, 32.2(2), 27.0(2), 26.1, 25.5(2). LCMS: m/e 320 (M-H)^“, ret time 2.21 min, column A, 4 minute gradient.

Intermediate 3

lH-Indole-6-carboxamide, 2-bromo-3-cyclohexyl-N-

[(dimethylamino)sulfonyl]-. l,l’-Carbonyldiimidazole (1.17 g, 7.2 mmol) was added to a stirred solution of 2-bromo-3-cyclohexyl-lH-indole-6-carboxylic acid (2.03 g, 6.3 mmol) in THF (6 mL) at 22 °C. The evolution of CO₂ was instantaneous and when it slowed the solution was heated at 50°C for 1 hr and then cooled to 22⁰C. N,N-Dimethylsulfamide (0.94 g, 7.56 mmol) was added followed by the dropwise addition of a solution of DBU (1.34 g ,8.8 mmol) in THF (4 mL). Stirring was continued for 24 hr. The mixture was partitioned between ethyl acetate and dilute HCl. The ethyl acetate layer was washed with water followed by brine and dried over Na₂SO₄. The extract was concentrated to dryness to leave the title product as a pale yellow friable foam, (2.0 g, 74 %, >90 % purity , estimated from NMR). ¹H NMR (300 MHz, DMSO-D6) δ ppm 1.28 – 1.49 (m, 3 H) 1.59 – 2.04 (m, 7 H) 2.74 – 2.82 (m, 1 H) 2.88 (s, 6 H) 7.57 (dd, J=8.42, 1.46 Hz, 1 H) 7.74 (d, J=8.78 Hz, 1 H) 7.91 (s, 1 H) 11.71 (s, 1 H) 12.08 (s, 1 H).

An alternative method for the preparation of lH-indole-6-carboxamide, 2- bromo-3-cyclohexyl-N-[(dimethylamino)sulfonyl]- is described below.

To a 1 L four necked round bottom flask equipped with a mechanical stirrer, a temperature controller, a N2 inlet , and a condenser, under N2, was added 2-bromo-3- cyclohexyl-lH-indole-6-carboxylic acid (102.0 g, 0.259 mol) and dry TΗF (300 mL). After stirring for 10 min, CDI (50.3 g, 0.31 mol) was added portion wise. The reaction mixture was then heated to 50 oC for 2 h. After cooling to 30 oC, N,N- dimethylaminosulfonamide (41.7 g, 0.336 mol) was added in one portion followed by addition of DBU (54.1 mL, 0.362 mol) drop wise over a period of 1 h. The reaction mixture was then stirred at rt for 20 h. The solvent was removed in vacuo and the residue was partitioned between EtOAc and 1 Ν HCl (1 : 1, 2 L). The organic layer was separated and the aqueous layer was extracted with EtOAc (500 mL). The combined organic layers were washed with brine (1.5 L) and dried over MgSO4. The solution was filtered and concentrated in vacuo to give the crude product (111.0 g). The crude product was suspended in EtOAc (400 mL) at 60 oC. To the suspension was added heptane (2 L) slowly. The resulting suspension was stirred and cooled to 0 oC. It was then filtered. The filter cake was rinsed with small amount of heptane and house vacuum air dried for 2 days. The product was collected as a white solid (92.0 g, 83%). ¹H ΝMR (MeOD, 300 MHz) δ 7.89 (s, H), 7.77 (d, J= 8.4 Hz, IH), 7.55 (dd, J= 8.4 and 1.8 Hz, IH), 3.01 (s, 6H), 2.73-2.95 (m, IH), 1.81-2.05 (m, 8H), 1.39-1.50 (m, 2H); m/z 429 (M +H)+. Intermediate 4

lH-Indole-6-carboxamide, 3-cyclohexyl-N-[(dimethylamino)sulfonyl]-2-(2- formyl-4-methoxyphenyl)-. A mixture of the 2-Bromo-3-cyclohexyl- N- [(dimethylamino)sulfonyl]-lH-indole-6-carboxamide (4.28g, 0.01 mol), 4-methoxy- 2-formylphenyl boronic acid (2.1%, 0.015 mol), 2-dicyclohexylphosphino-2′,6′- dimethoxy-biphenyl (41 mg, 0.0001 mol), palladium acetate (11.2 mg), and finely ground potassium carbonate (4.24g, 0.02 mol) in toluene (30 mL) was stirred under reflux and under nitrogen for 30 min, at which time LC/MS analysis showed the reaction to be complete. The reaction mixture was then diluted with ethyl acetate and water, and then acidified with an excess of dilute HCl. The ethyl acetate layer was then collected and washed with dilute HCl, water and brine. The organic solution was then dried (magnesium sulfate), filtered and concentrated to give a gum. The gum was diluted with hexanes (250 ml) and ethyl acetate (25 mL), and the mixture was stirred for 20 hr at 22° C during which time the product was transformed into a bright yellow granular solid (4.8 g) which was used directly without further purification.

An alternative procedure for the preparation of lH-indole-6-carboxamide, 3- cyclohexyl-N-[(dimethylamino)sulfonyl]-2-(2-formyl-4-methoxyphenyl)- is provided below:

To a slurried solution of 2-bromo-3-cyclohexyl-N-[(dimethylamino)sulfonyl]- indole-6-carboxamide (54.0 g, 126 mmol), 4-methoxy-2-formylphenylboronic acid (29.5 g, 164 mmol) and LiCl (13.3 g, 315 mmol) in EtOH/toluene (1 : 1, 1 L) was added a solution of Na₂CO3 (40.1 g, 379 mmol) in water (380 mL). The reaction mixture was stirred 10 min. and then Pd(PPh3)4 (11.3 g, 10.0 mmol) was added. The reaction solution was flushed with nitrogen and heated at 70 °C (internal monitoring) overnight and then cooled to rt. The reaction was diluted with EtOAc (1 L) and EtOH (100 mL), washed carefully with IN aqueous HCl (1 L) and brine (500 mL), dried (MgSO4), filtered and concentrated. The residual solids were stirred with Et20 (600 mL) for Ih and collected by filtration to yield lH-indole-6-carboxamide, 3- cyclohexyl-N-[(dimethylamino)sulfonyl]-2-(2-formyl-4-methoxyphenyl)- (52.8g, 109 mmol, 87%) as a yellow powder which was used without further purification. IHNMR (300 MHz, d6-DMSO) δ 11.66 (s, IH), 8.17 (s, IH), 7.75 (d, J = 8.4 Hz, IH), 7.74 (d, J = 8.4 Hz, IH), 7.59 (dd, J = 1.4, 8.4 Hz, IH), 7.23 – 7.16 (m, 2H), 7.08 (dd, J = 2.6, 8.4 Hz, IH), 6.54 (d, J = 8.8 Hz, IH), 3.86 (s, 3H), 3.22 – 3.08 (m, IH), 2.91 (s, 6H), 2.00 – 1.74 (m, 7H), 1.60 – 1.38 (m, 3H). 13CNMR (75 MHz, CDC13) δ 165.7, 158.8, 147.2, 139.1, 134.3, 132.0, 123.4, 122.0, 119.2, 118.2, 114.8, 112.3, 110.4, 109.8, 79.6, 45.9, 37.2(2), 34.7, 32.0(2), 25.9(2), 24.9. LCMS: m/e 482 (M- H)^“, ret time 2.56 min, column A, 4 minute gradient.

Intermediate 5

6H-Isoindolo[2,l-a]indole-3-carboxamide, 11-cyclohexyl-N-

[(dimethylamino)sulfonyl]-6-ethoxy-8-methoxy-. To a 5 L four necked round bottom flask equipped with a temperature controller, a condenser, a N2 inlet and a mechanical stirrer, was charged toluene (900 mL), EtOH (900 mL), 2-bromo-3- cyclohexyl-N^NjN-dimethylsulfamoyiyiH-indole-ό-carboxamide (90 g, 0.21 mol), 2-formyl-4-methoxyphenylboronic acid (49.2 g, 0.273 mol) and LiCl (22.1 g, 0.525 mol). The resulting solution was bubbled with Ν₂ for 15 mins. A solution of Na₂CO3 (66.8 g, 0.63 mol) in Η₂O (675 mL) was added and the reaction mixture was bubbled with N₂ for another (10 mins). Pd(PPh₃)₄ (7.0 g, 6.3 mmol) was added and the reaction mixture was heated to 70 °C for 20 h. After cooling to 35 °C, a solution of 1 N HCl (1.5 L) was added slowly. The resulting mixture was transferred to a 6 L separatory funnel and extracted with EtOAc (2 X 1.5 L). The combined organic extracts were washed with brine (2 L), dried over MgSO4, filtered and concentrated in vacuo to give a yellow solid, which was triturated with 20% EtOAc in hexane (450 mL, 50 °C to 0 °C) to give 3-cyclohexyl-N-(N,N-dimethylsulfamoyl)-2-(2-formyl-4- methoxyphenyl)-lH-indole-6-carboxamide(65.9 g) as a yellow solid. HPLC purity, 98%.

The mother liquid from the trituration was concentrated in vacuo. The residue was refluxed with EtOH (50 mL) for 3 h. The solution was then cooled to 0 °C. The precipitates were filtered and washed with cooled TBME (5 °C) (20 mL). The filter cake was house vacuum air dried to give a further quantity of the title compound as a white solid (16.0 g). HPLC purity, 99%. ¹H NMR (CDC13, 300 MHz) δ 8.75 (s, IH), 7.96 (s, IH), 7.73 (d, J= 8.4 Hz, IH), 7.67 (d, J= 8.4 Hz, IH), 7.45 (dd, J= 8.4 and 1.4 Hz, IH), 7.09 (d, J= 2.2 Hz, IH), 6.98 (dd, J= 8.4 and 2.2 Hz, IH), 6.50 (s, IH), 3.86 (s, 3H), 3.05 (s, 6H), 2.92-3.13 (m, 3H), 1.85-1.93 (m, 7 H), 1.40-1.42 (m, 3H), 1.05 (t, J= 7.1 Hz, 3H). m/z 512 (M + H)+.

Intermediate 6

lH-indole-6-carboxamide, 3-cyclohexyl-N-[(dimethylamino)sulfonyl]-2-(2- formyl-4-methoxyphenyl)-. 1 l-cyclohexyl-N-(N,N-dimethylsulfamoyl)-6-ethoxy-8- methoxy-6H-isoindolo[2,l-a]indole-3-carboxamide was dissolved in THF (75 mL). To the solution was added a solution of 2 N HCl (300 mL). The mixture was vigorously stirred under N2 at rt for 16 h. The resulting suspension was filtered and washed with cooled TBME (2 X 30 mL). the filer cake was vacuum air dried overnight to give the title compound as a yellow solid. HPLC purity, 99% ¹H NMR (DMSO-d6, 300 MHz) δ 11.65 (s, IH), 8.16 (s, IH), 7.76 (d, J= 5.9 Hz, IH), 7.73 (d, J= 5.9 Hz, IH), 7.58 (dd, J= 8.5 and 1.5 Hz, IH), 7.17-7.20 (m, 2H), 7.08 (dd, J = 8.5 and 1.4 Hz, IH), 6.55 (d, J= 8.6 Hz, IH), 3.86 (s, 3H), 3.14-3.18 (m, IH), 2.91 (s, 6H), 1.75-1.99 (m, 7H), 1.48-1.60 (m, 3H); m/z 484 (M + H)+.

Intermediate 7

7H-Indolo[2, 1-a] ‘ [2] benzazepine-6-carboxylic acid, 13-cyclohexyl-10- [[[(dimethylamino)sulfonyl] amino] carbonyl]-3-methoxy-, methyl ester. A mixture of the 3-cyclohexyl-N-(N,N-dimethylsulfamoyl)-2-(2-formyl-4-methoxyphenyl)-lH- indole-6-carboxamide (4.8g, 0.01 mol), methyl 2-(dimethoxyphosphoryl)acrylate (9.7 g, 0.02 mol) and cesium carbonate (7.1g, 0.02 mol) in DMF (28mL) was stirred for 20 hr at an oil bath temperature of 55 ° C. The mixture was poured into ice-water and acidified with dilute HCl to precipitate the crude product. The solid was collected, dried and flash chromatographed on Siθ₂ (11Og) using an ethyl acetate and methylene chloride (1: 10) solution containing 2% acetic acid. Homogeneous fractions were combined and evaporated to afford the title compound as a pale yellow solid (3.9g, 71 % yield). MS: 552 (M=H+).

An alternate procedure for the preparation of 7H-indolo[2,l- a] [2]benzazepine-6-carboxylic acid, 13-cyclohexyl-10- [[[(dimethylamino)sulfonyl]amino]carbonyl]-3-methoxy-, methyl ester is provided below. A solution of l l-cyclohexyl-N-[(dimethylamino)sulfonyl]-6-hydroxy-8- methoxy-6H-isoindolo[2,l-a]indole-3-carboxamide (cyclic hemiaminal) (63.0 g, 130 mmol), methyl 2-(dimethoxyphosphoryl)acrylate (60 g, 261 mmol), cesium carbonate (106 g, 326 mmol) in DMF (400 mL) was heated at 60 °C (bath temp) for 4.5h. Additional methyl 2-(dimethoxyphosphoryl)acrylate (15 g, 65 mmol) and cesium carbonate (21.2 g, 65 mmol) were added and the reaction was heated at 60 °C overnight then and cooled to rt. The stirring reaction mixture was diluted with H₂O (1 L), slowly neutralized with IN aqueous HCl (800 mL), stirred 3h, and then the precipitates were collected by filtration. The solids were triturated with Et20 (800 mL) and dried to yield methyl 7H-indolo[2,l-a][2]benzazepine-6-carboxylic acid, 13- cyclohexyl-10-[[[(dimethylamino)sulfonyl]amino]carbonyl]-3-methoxy-, methyl ester (70.2 g, 127 mmol, 98%) as a yellow solid which was used without further purification. IHNMR (300 MHz, CDC13) δ 8.67 (s, IH), 8.09 (s, IH), 7.86 (d, J = 8.4 Hz, IH), 7.80 (s, IH), 7.50 (d, J = 8.4 Hz, IH), 7.42 (d, J = 8.8 Hz, IH), 7.08 (dd, J = 2.6, 8.8 Hz, IH), 6.98 (d, J = 2.6 Hz, IH), 5.75 – 5.51 (m, IH), 4.29 – 4.01 (m, IH), 3.89 (s, 3H), 3.82 (s, 3H), 3.05 (s, 6H), 2.87 – 2.73 (m, IH), 2.11 – 1.12 (m, 10H). LCMS: m/e 550 (M-H)-, ret time 3.21 min, column A, 4 minute gradient.

Example 1

Cycloprop[d]indolo[2,l-a] [2]benzazepine-5-carboxamide, 8-cyclohexyl-N- [(dimethylamino)sulfonyl]-l,la,2,12b-tetrahydro-ll-methoxy-la-[(3-methyl-3,8- diazabicyclo[3.2.1]oct-8-yl)carbonyl]-, (+/-)-. TBTU (43.7 mg, 0.136mmol) and DIPEA (0.095 mL, 0.544 mmol) were added to a solution of (+/-) cycloprop[d]indolo[2,l-a][2]benzazepine-la(2H)-carboxylic acid, 8-cyclohexyl-5- [[[(dimethylamino)sulfonyl]amino]carbonyl]-l,12b-dihydro-l 1-methoxy- (50 mg, 0.0906 mmol) in DMSO (2.0 mL). The reaction mixture was stirred at rt for 15 min. 3-Methyl-3,8-diaza-bicyclo[3.2. l]octane dihydrochloride {J & W PharmLab, LLC Morrisville, PA 19067-3620}. (27.1 mg, 0. 136 mmol) was then added and the reaction mixture was stirred at rt for 3 hr. It was then concentrated and the residue was purified by preparative reverse phase HPLC to give the final product as a yellow solid, (32 mg, 46% yield). MS m/z 660(MH⁺), Retention time: 2.445 min IH NMR (300 MHz, MeOD) δ ppm 0.20 (m, 0.23 H) 1.11 – 2.25 (m, 15.77 H) 2.58 (m, 0.23 H) 2.69 (m, 0.77 H) 2.75 – 3.11 (m, 10 H) 3.28 – 3.75 (m, 5 H) 3.91 (s, 2.31 H) 3.92 (s, 0.69 H) 4.15 – 4.37 (m, 1 H) 4.68 (m ,br, 1 H) 4.94 – 5.00 (m, 0.23 H) 5.16 (d, J=15.00 Hz, 0.77 H) 7.00 – 7.09 (m, 1 H) 7.18 (d, J=2.56 Hz, 0.23 H) 7.21 (d, J=2.56 Hz, 0.77 H) 7.33 (d, J=8.41 Hz, 0.77 H) 7.35 (d, J=8.42 Hz, 0.23 H) 7.57 (dd, J=8.42, 1.46 Hz, 0.77 H) 7.62 (dd, J=8.78, 1.46 Hz, 0.23 H) 7.91 (d, J=8.42 Hz, 0.77 H) 7.93 (d, J=8.42 Hz, 0.23 H) 8.00 (s, 0.77 H) 8.07 (s, 0.23 H).

Example 4

Cycloprop[d]indolo[2,l-a] [2]benzazepine-5-carboxamide, 8-cyclohexyl-N- [(dimethylamino)sulfonylj '- 1 , Ia, 2, 12b-tetrahydro-ll-methoxy-la-[(8-methyl-3, 8- diazabicyclo[3.2.1]oct-3-yl)carbonyl]-, (+/-)-. To a solution of (+/-) cycloprop[d]indolo[2,l-a][2]benzazepine-5-carboxamide, 8-cyclohexyl-la-(3,8- diazabicyclo[3.2.1]oct-3-ylcarbonyl)-N-[(dimethylamino)sulfonyl]-l,la,2,12b- tetrahydro-11-methoxy- (54 mg, 0.071 mmol) in methanol (3 mL), paraformaldehyde (6.4 mg, 0.213 mmol), ZnCl₂ (29 mg, 0.213 mmol) and

Na(CN)BH₃ (13.4 mg, 0.213 mmol) were added. The resultant mixture was heated at 60°C for 2hr, and then cooled to rt. The solid present was removed by filtration, and the filtrate was concentrated under vacuum and the residue purified by preparative reverse phase HPLC to give the title compound as a light yellow colored solid, (37 mg, 67% yield). MS ml 660(MH⁺), Retention time: 2.495 min. IH NMR (500 MHz, MeOD) δ ppm 0.21 (m, 0.3 H) 1.13 (m, 0.3 H) 1.18 – 2.22 (m, 15.4 H) 2.58 (m, 0.3 H) 2.68 (m, 0.7 H) 2.76 – 3.11 (m, 11 H) 3.32 – 3.37 (m, 1 H) 3.63 (d, J=15.56 Hz, 0.7 H) 3.82 – 4.32 (m, 7.3 H) 4.88 – 4.92 (m, 0.3 H) 5.08 (d, J=15.56 Hz, 0.7 H) 7.00 – 7.08 (m, 1 H) 7.18 (d, J=2.14 Hz, 0.3 H) 7.21 (d, J=2.14 Hz, 0.7 H) 7.32 (d, J=8.55 Hz, 0.7 H) 7.35 (d, J=8.55 Hz, 0.3H) 7.57 (d, J=7.93 Hz, 0.7 H) 7.62 (dd, J=8.39, 1.37 Hz, 0.3 H) 7.91 (d, J=8.55 Hz, 0.7 H) 7.93 – 7.99 (m, 1 H) 8.09 (s, 0.3 H).

Example 6

Cycloprop [d] indolo [2, 1 -a] [2]benzazepine-5-carboxamide, 8-cyclohexyl-N- [(dimethylamino)sulfonyl]-l,la,2,12b-tetrahydro-ll-methoxy-la-[(3-methyl-3,8- diazabicyclo[3.2.1]oct-8-yl)carbonyl]-, (laR,12bS)-. To a solution of (-) cycloprop[d]indolo[2,l-a][2]benzazepine-la(2H)-carboxylic acid, 8-cyclohexyl-5- [[[(dimethylamino)sulfonyl]amino]carbonyl]-l,12b-dihydro-l 1-methoxy- (204 mg, 0.37 mmol) in DMSO (8.0 mL), TBTU (178 mg, 0.555 mmol) and DIPEA (0.39 mL, 2.22 mmol) were added. The reaction mixture was stirred at rt for 15 min. Then 3- methyl-3,8-diaza-bicyclo[3.2.1]octane dihydrochloride (111 mg, 0. 555 mmol) was added and the reaction mixture was stirred at rt for 2 hr. It was then concentrated and the residue was purified by preparative reverse phase HPLC to give a yellow solid as final TFA salt. (265 mg, 92% yield). Average Specific Rotation: -53.56° Solvent, MeOH.; Wavelength 589 nm; 50 cm cell. MS m/z 660(MH⁺), Retention time: 3.035 min. ¹H NMR (300 MHz, MeOD) δ ppm 0.20 (m, 0.23 H) 1.11 – 2.25 (m, 15.77 H) 2.58 (m, 0.23 H) 2.69 (m, 0.77 H) 2.75 – 3.11 (m, 10 H) 3.28 – 3.75 (m, 5 H) 3.91 (s, 2.31 H) 3.92 (s, 0.69 H) 4.15 – 4.37 (m, 1 H) 4.68 (m ,br, 1 H) 4.94 – 5.00 (m, 0.23 H) 5.16 (d, J=15.00 Hz, 0.77 H) 7.00 – 7.09 (m, 1 H) 7.18 (d, J=2.56 Hz, 0.23 H) 7.21 (d, J=2.56 Hz, 0.77 H) 7.33 (d, J=8.41 Hz, 0.77 H) 7.35 (d, J=8.42 Hz, 0.23 H) 7.57 (dd, J=8.42, 1.46 Hz, 0.77 H) 7.62 (dd, J=8.78, 1.46 Hz, 0.23 H) 7.91 (d, J=8.42 Hz, 0.77 H) 7.93 (d, J=8.42 Hz, 0.23 H) 8.00 (s, 0.77 H) 8.07 (s, 0.23 H). An alternate procedure for the synthesis of cycloprop[d]indolo[2,l- a][2]benzazepine-5-carboxamide, 8-cyclohexyl-N-[(dimethylamino)sulfonyl]- l,la,2,12b-tetrahydro-l l-methoxy-la-[(3-methyl-3,8-diazabicyclo[3.2.1]oct-8- yl)carbonyl]-, (laR,12bS)-rel-(-)-is provided below. To a mixture of (-) cycloprop[<i]indolo[2,l-α][2]benzazepine-la(2H)-carboxylic acid, 8-cyclohexyl-5- [[[(dimethylamino)sulfonyl]amino]carbonyl]-l,12b-dihydro-l 1-methoxy- (25.2 g, 45.68 mmol) and 3-methyl-3,8-diazabicyclo-[3.2.1]octane dihydrochloride (10.0 g, 50.22 mmol) in anhydrous MeCN (300 mL) was added DIPEA (23.62 g, 182.72 mmol) under N₂. After 15 min, TBTU (16.12 g, 50.22 mmol) was added. The reaction solution was stirred for 30 min under N₂. The ΗPLC indicated the disappearance of starting material. The solvent in the solution was evaporated to give a foam. This was dissolved in EtOAc (2.5 L), washed with H₂O (1.5 L), H₂O/brine (8:2) (1.5 L), brine (1.5 L), dried over Na₂SO₄ and evaporated to give 28.8 g of crude product. This solid was pooled with 45.4 g of material obtained from five separated reactions to afford a total of 74.2 g of crude product. This was passed through a pad of silica gel (E. Merck 230-400 mesh, 1 kg), eluting with MeOH/CH₂Cl₂ (2.5:97.5). After evaporation, it gave a foam, which was treated with EtOAc and hexane to turn into a solid. After drying at 50 °C under vacuum for 7 h, the GC analysis indicated it has 1.4% each of EtOAc and hexane. After further drying at 61-64 °C, the GC analysis indicated it still has 1.0% of hexane and 1.4% of EtOAc. The product was dissolved in Et₂O and slowly evaporated in vacuum three times, dried at 60 °C under vacuum for 3 h to give 68.3 g. This was washed with H₂O (900 mL) and redried at 68 °C under vacuum for 7 h to give 67.1 g (77% yield) of the compound of example 6. The GC analysis indicated it has 0.97% Of Et₂O. HPLC conditions column: Cadenza CD-C18 3 x 250 mm; UV: 257 and 220 nm; 25 °C; flow rate: 0.5 mL/min; gradient time: 38 min, 0 – 80% B (0 – 35 min) and 80% B (35 – 38 min); solvent A: 25 nM CH₃COONH₄ at pH 4.7 in water, solvent B: MeCN. HPLC purity 99.7% (Rt 26.54 min); Chiral HPLC conditions column: Regis (S₅S) Whelk-Ol 250 x 4.6 mm; UV 258nm; 35 °C; flow rate 2.0 mL/min; mobile phase C0₂/Me0H; gradient time 20 min, 30% MeOH (0 – 1 min), 30 – 48% MeOH (1 – 19 min), 48% MeOH (19 – 20 min). Chiral HPLC purity > 99.8% (Rt 16.60 min); LC/MS (ES⁺) 660.36 (M+H, 100); HRMS: calcd. 660.3220, found 660.3197; [α]_D ^{25 C} – 79.66 ° (c 1.06, MeOH); Anal. Calcd for C₃₆H₄₅N₅O₅S-O-O H₂O»0.09 Et₂O: C, 64.53; H, 7.00; N, 10.35; S, 4.74; H₂O, 1.51; Et₂O, 0.97. Found: C, 64.50; H, 7.12; N, 10.41; S, 5.14; H₂O, 1.52; Et₂O, 0.97. The absolute stereochemistry of cycloprop[d]indolo[2,l- a][2]benzazepine-5-carboxamide, 8-cyclohexyl-N-[(dimethylamino)sulfonyl]- l,la,2,12b-tetrahydro-l l-methoxy-la-[(3-methyl-3,8-diazabicyclo[3.2.1]oct-8- yl)carbonyl]-, (laR,12bS)-rel-(-)- is as drawn above, and was determined from an x- ray crystal structure obtained on the (R)-camphorsulfonic acid salt.

Additionally, the following salts were prepared: hydrochloride, phosphate, acetate, sulfate, camsylate, sodium, calcium, and magnesium. The hydrochloride salt had the following characteristics. DSC: small, broad endotherm from 25°C to 75°C, and potential melt/degradation endotherm with peak at temperatures ranging between 253 °C and 258 °C; TGA: Early weight loss from 25°C to 75°C ranging between 0.003% and 1.5%, and degradation weight loss starting at approximately 200°C.