[F-18](2S,4S)-4-(3-Fluoropropyl)glutamine

Uncategorized Comments Off

May 302016

[F-18](2S,4S)-4-(3-Fluoropropyl)glutamine

CAS 1196963-79-7

MF C₈ H₁₅ F N₂ O₃

Heptanoic acid, 2-amino-4-(aminocarbonyl)-7-(fluoro-¹⁸F)-, (2S,4S)-


[¹⁸F](2S,4S)-4-FPGln	[¹⁸F](2S,4S)-4-(3-fluoropropyl)glutamine, 4

The early diagnosis of malignant tumors plays a very important role in the survival prognosis of cancer patients. In this non-invasive diagnosis, diagnostic imaging procedures are an important tool. In the last few years has mainly PET technology (P ositronen- E mission- Tomographie) proved to be particularly useful. The sensitivity and specificity of PET technology depends significantly on the used signal-emitting substance (tracer) and their distribution in the body from. In the search for suitable tracers one tries to take advantage of certain properties of tumors differ, the tumor tissue from healthy, surrounding tissue. The preferred commercially used isotope which finds application for PET, ¹⁸ F ¹⁸ F represents by its short half-life of less than 2 hours special requirements for the preparation of suitable tracer. Complex, long synthetic routes and purifications are with this isotope is not possible, because otherwise a significant portion of the radioactivity of the isotope has already decayed before the tracer can be used for diagnosis. It is therefore often not possible to established synthetic routes for non-radioactive fluorination to be applied to the synthesis of¹⁸ F-tracer. Furthermore, the high specific activity of ¹⁸ F (80 GBq / nmol) at very low substance amounts of ^[18 F] fluoride for the tracer synthesis, which in turn an extreme excess of precursor-related and the success of a non-radioactive fluorination based Radio synthetic strategy designed unpredictable

FDG ^([18 F] F 2 luoro d esoxy lukose g) -PET is a widely accepted and popular tool in the diagnosis and other clinical tracking of tumor diseases. Malignant tumors compete with the host organism to glucose supply to the nutrient supply (Warburg O. About the metabolism of carcinoma cell Biochem;.. Kellof G. Progress and Promise of FDG PET Imaging for Cancer Patient Management and Oncologic Drug Development Clin Cancer Res 2005;.. 11 (8): 2785-2807) where tumor cells compared to surrounding cells of normal tissue usually an increased glucose metabolism. This is used when using fluorodeoxyglucose (FDG), a glucose derivative, which is amplified transported into the cells, but there included metabolically after phosphorylation as FDG-6-phosphate (“Warburg effect”). ¹⁸ F-labeled FDG is Therefore, an effective tracer for the detection of tumors in patients using PET technology. Imaging were looking for new PET tracers in recent years increasingly amino acids for ¹⁸ F PET used (eg (review): Eur J Nucl Med Mol Imaging 2002 May; 29 (5):.. 681-90). In this case, some of the ¹⁸ F-labeled amino acids for the measurement of the speed rate of protein synthesis, the most useful derivatives but for the direct measurement of the cellular uptake in the tumor. Known ¹⁸ F-labeled amino acids are, for example, from tyrosine, phenylalanine, proline, aspartic and unnatural amino acids derived (eg J. Nucl Med 1991; 32:.. 1338-1346, J Nucl Med 1996; 37: 320-325, J Nucl Med 2001; 42: 752-754 J Nucl Med and 1999, 40: 331-338).. Glutamic acid and glutamine than ¹⁸ F-labeled derivatives not known, whereas non-radioactive fluorinated glutamine and glutamic acid derivatives are well known; Thus, for Example those which at γ-position (for Ex (review):Amino Acids (2003) April; 24 (3):… 245-61).. or at β-position (e.g. ExTetrahedron. Lett. .; 30; 14; 1989, 1799-1802, J. Org Chem .; 54; 2; 1989, 498-500, Tetrahedron: Asymmetry, 12, 9; 2001; 1303-1312) havefluorine..

Of glutamic acid having the chemical functionalities protecting groups in β and γ position or a leaving group, has already been reported in the past. So was informed of glutamate as mesylate or bromide in γ-position whose acid and amine functions were provided with ester or Z-protecting groups (J. Chem Soc Perkin Trans. 1;.. 1986, 1323-1328) or, for example, of γ-chloro-glutamic acid without protecting groups(Synthesis, (1973); 44-46). About similar derivatives, but where the leaving group is positioned in β-position has also been reported on several occasions. Z Ex. Chem. Pharm. Bull .; 17; 5; (1969); 879-885,J.Gen.Chem.USSR (Engl.Transl.); 38; (1968); 1645-1648, Tetrahedron Lett .; 27; 19; (1986); 2143-2144, Chem. Pharm. Bull .; EN; 17; 5; 1969;873-878, patent FR 1461184 , Patent JP 13142 .)

The current PET tracers, which are used for tumor diagnosis have some undisputed disadvantages: in FDG accumulates preferably in those cells with increased glucose metabolism on, but there are also other pathological and physiological conditions of increased glucose metabolism in the cells involved and tissues, eg, Ex. of infection or wound healing (summarized in J. Nucl. Med. Technol. (2005), 33, 145-155). It is still often difficult to decide whether a detected by FDG-PET lesion actually neoplastic origin or due to other physiological or pathological state of the tissue. Overall, the diagnostic activity by FDG-PET in oncology has a sensitivity of 84% and a specificity of 88% to(Gambhir et al., ” A tabulated summary of the FDG PET literature “J. Nucl. Med. 2001, 42, 1- 93S). Tumors in the brain can be represented very difficult in healthy brain tissue, for example, by the high accumulation of FDG.

The previously known ¹⁸ F-labeled amino acid derivatives are in some cases well suited to detect tumors in the brain ((review): Eur J Nucl Med Mol Imaging 2002 May; 29 (5):. 681-90), but they can in other tumors do not compete with the imaging properties of the “gold standard” ^[18 F] 2-FDG. The metabolic accumulation and retention of previously F-18 labeled amino acids in tumorous tissue is usually lower than for FDG. Moreover, the accessibility of isomerically pure F-18-labeled non-aromatic amino acids is chemically very demanding.

Similar to glucose increased metabolism in proliferating tumor cells has been described (Medina, J Nutr 1131: 2539S-2542S, 2001; Souba, Ann Surg 218:. 715-728, 1993) for glutamic acid and glutamine. The increased rate of protein and nucleic acid synthesis and energy production per se be accepted as reasons for increased Glutaminkonsum of tumor cells. The synthesis of the corresponding C-11 and C-14 labeled with the natural substrate thus identical compounds, has already been described in the literature (eg. Ex.Antoni, enzymes Catalyzed Synthesis of L- [4-C-11] Aspartate and L – [5-C-11] Glutamate J. Labelled Compd Radiopharm 44; (4) 2001: 287-294) and Buchanan, The biosynthesis of showdomycin: studies with stable isotopes and the determination of principal precursor J….. Chem. Soc. Chem. Commun .; EN; 22; 1984, 1515-1517). First indications with the C-11 labeled compound indicate no significant tumor accumulation.

Although the growth and proliferation of most tumors is fueled by glucose, some tumors are more likely to metabolize glutamine. In particular, tumor cells with the upregulated c-Myc gene are generally reprogrammed to utilize glutamine. We have developed new 3-fluoropropyl analogs of glutamine, namely [(18)F](2S,4R)- and [(18)F](2S,4S)-4-(3-fluoropropyl)glutamine, 3 and 4, to be used as probes for studying glutamine metabolism in these tumor cells. Optically pure isomers labeled with (18)F and (19)F (2S,4S) and (2S,4R)-4-(3-fluoropropyl)glutamine were synthesized via different routes and isolated in high radiochemical purity (≥95%). Cell uptake studies of both isomers showed that they were taken up efficiently by 9L tumor cells with a steady increase over a time frame of 120 min. At 120 min, their uptake was approximately two times higher than that of l-[(3)H]glutamine ([(3)H]Gln). These in vitro cell uptake studies suggested that the new probes are potential tumor imaging agents. Yet, the lower chemical yield of the precursor for 3, as well as the low radiochemical yield for 3, limits the availability of [(18)F](2S,4R)-4-(3-fluoropropyl)glutamine, 3. We, therefore, focused on [(18)F](2S,4S)-4-(3-fluoropropyl)glutamine, 4. The in vitro cell uptake studies suggested that the new probe, [(18)F](2S,4S)-4-(3-fluoropropyl)glutamine, 4, is most sensitive to the LAT transport system, followed by System N and ASC transporters. A dual-isotope experiment using l-[(3)H]glutamine and the new probe showed that the uptake of [(3)H]Gln into 9L cells was highly associated with macromolecules (>90%), whereas the [(18)F](2S,4S)-4-(3-fluoropropyl)glutamine, 4, was not (<10%). This suggests a different mechanism of retention. In vivo PET imaging studies demonstrated tumor-specific uptake in rats bearing 9L xenographs with an excellent tumor to muscle ratio (maximum of ∼8 at 40 min). [(18)F](2S,4S)-4-(3-fluoropropyl)glutamine, 4, may be useful for testing tumors that may metabolize glutamine related amino acids.

[¹⁸F](2S,4S)-4-(3-Fluoropropyl)glutamine as a Tumor Imaging Agent

http://pubs.acs.org/doi/full/10.1021/mp500236y

Zehui Wu†, Zhihao Zha†, Genxun Li†, Brian P. Lieberman†, Seok Rye Choi†, Karl Ploessl†, and Hank F. Kung^*†‡

^†Departments of Radiology and ^‡Pharmacology, University of Pennsylvania, 3700 Market Street, Philadelphia, Pennsylvania 19104, United States

Mol. Pharmaceutics, 2014, 11 (11), pp 3852–3866

DOI: 10.1021/mp500236y

Publication Date (Web): August 05, 2014

*Email: kunghf@sunmac.spect.upenn.edu. Phone: 215-662-3096. Fax: 215-349-5035.

ACS AuthorChoice – 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.

This article is part of the Positron Emission Tomography: State of the Art special issue.

Abstract

Although the growth and proliferation of most tumors is fueled by glucose, some tumors are more likely to metabolize glutamine. In particular, tumor cells with the upregulated c-Myc gene are generally reprogrammed to utilize glutamine. We have developed new 3-fluoropropyl analogs of glutamine, namely [¹⁸F](2S,4R)- and [¹⁸F](2S,4S)-4-(3-fluoropropyl)glutamine, 3 and 4, to be used as probes for studying glutamine metabolism in these tumor cells. Optically pure isomers labeled with ¹⁸F and ¹⁹F (2S,4S) and (2S,4R)-4-(3-fluoropropyl)glutamine were synthesized via different routes and isolated in high radiochemical purity (≥95%). Cell uptake studies of both isomers showed that they were taken up efficiently by 9L tumor cells with a steady increase over a time frame of 120 min. At 120 min, their uptake was approximately two times higher than that of l-[³H]glutamine ([³H]Gln). These in vitro cell uptake studies suggested that the new probes are potential tumor imaging agents. Yet, the lower chemical yield of the precursor for 3, as well as the low radiochemical yield for 3, limits the availability of [¹⁸F](2S,4R)-4-(3-fluoropropyl)glutamine, 3. We, therefore, focused on [¹⁸F](2S,4S)-4-(3-fluoropropyl)glutamine, 4. The in vitro cell uptake studies suggested that the new probe, [¹⁸F](2S,4S)-4-(3-fluoropropyl)glutamine, 4, is most sensitive to the LAT transport system, followed by System N and ASC transporters. A dual-isotope experiment using l-[³H]glutamine and the new probe showed that the uptake of [³H]Gln into 9L cells was highly associated with macromolecules (>90%), whereas the [¹⁸F](2S,4S)-4-(3-fluoropropyl)glutamine, 4, was not (<10%). This suggests a different mechanism of retention. In vivo PET imaging studies demonstrated tumor-specific uptake in rats bearing 9L xenographs with an excellent tumor to muscle ratio (maximum of ∼8 at 40 min). [¹⁸F](2S,4S)-4-(3-fluoropropyl)glutamine, 4, may be useful for testing tumors that may metabolize glutamine related amino acids.

PATENT

US 20100290991

http://www.google.com/patents/US20100290991

PATENT

WO 2009141091

https://patentscope.wipo.int/search/ko/detail.jsf?docId=WO2009141091&recNum=70&maxRec=287&office=&prevFilter=%26fq%3DOF%3ACU&sortOption=Relevance&queryString=&tab=PCTDescription

PATENT

http://www.google.co.ug/patents/EP2123621A1?cl=en

REFERENCES

https://www.researchgate.net/publication/264538736_F-182S4S-4-3-Fluoropropylglutamine_as_a_Tumor_Imaging_Agent

Molecular Pharmaceutics (2014), 11(11), 3852-3866

EP1923382A1 *	18 Nov 2006	21 May 2008	Bayer Schering Pharma Aktiengesellschaft	[18F] labelled L-glutamic acid, [18F] labelled glutamine, their derivatives, their use and processes for their preparation
FR1461184A				Title not available
JPS58113142A				Title not available
WO2008052788A1 *	30 Oct 2007	8 May 2008	Bayer Schering Pharma Aktiengesellschaft	[f-18]-labeled l-glutamic acid, [f-18]-labeled l-glutamine, derivatives thereof and use thereof and processes for their preparation

////////

FDA approves new diagnostic imaging agent FLUCICLOVINE F-18 to detect recurrent prostate cancer

FDA 2016, Uncategorized Comments Off

May 282016

FLUCICLOVINE F-18

Cyclobutanecarboxylic acid, 1-amino-3-(fluoro-¹⁸F)-, trans- [

Molecular FormulaC₅H₈¹⁸FNO₂
Average mass132.124 Da

Axumin (fluciclovine F 18)

fluciclovinum (¹⁸F)

GE-148

NMK36

trans-1-Amino-3-(¹⁸F)fluorcyclobutancarbonsäure [German] [ACD/IUPAC Name]

trans-1-Amino-3-(¹⁸F)fluorocyclobutanecarboxylic acid [ACD/IUPAC Name]

UNII-38R1Q0L1ZE

anti-1-amino-3-[18F]fluorocyclobutane-1-carboxylic acid

cas 222727-39-1

FDA approves new diagnostic imaging agent to detect recurrent prostate cancer

05/27/2016 11:27 AM EDT

The U.S. Food and Drug Administration today approved Axumin, a radioactive diagnostic agent for injection. Axumin is indicated for positron emission tomography (PET) imaging in men with suspected prostate cancer recurrence based on elevated prostate specific antigen (PSA) levels following prior treatment.

May 27, 2016

Release

Prostate cancer is the second leading cause of death from cancer in U.S. men. In patients with suspected cancer recurrence after primary treatment, accurate staging is an important objective in improving management and outcomes.

“Imaging tests are not able to determine the location of the recurrent prostate cancer when the PSA is at very low levels,” said Libero Marzella, M.D., Ph.D., director of the Division of Medical Imaging Products in the FDA’s Center for Drug Evaluation and Research. “Axumin is shown to provide another accurate imaging approach for these patients.”

Two studies evaluated the safety and efficacy of Axumin for imaging prostate cancer in patients with recurrent disease. The first compared 105 Axumin scans in men with suspected recurrence of prostate cancer to the histopathology (the study of tissue changes caused by disease) obtained by prostate biopsy and by biopsies of suspicious imaged lesions. Radiologists onsite read the scans initially; subsequently, three independent radiologists read the same scans in a blinded study.

The second study evaluated the agreement between 96 Axumin and C11 choline (an approved PET scan imaging test) scans in patients with median PSA values of 1.44 ng/mL. Radiologists on-site read the scans, and the same three independent radiologists who read the scans in the first study read the Axumin scans in this second blinded study. The results of the independent scan readings were generally consistent with one another, and confirmed the results of the onsite scan readings. Both studies supported the safety and efficacy of Axumin for imaging prostate cancer in men with elevated PSA levels following prior treatment.

Axumin is a radioactive drug and should be handled with appropriate safety measures to minimize radiation exposure to patients and healthcare providers during administration. Image interpretation errors can occur with Axumin PET imaging. A negative image does not rule out the presence of recurrent prostate cancer and a positive image does not confirm the presence of recurrent prostate cancer. Clinical correlation, which may include histopathological evaluation of the suspected recurrence site, is recommended.

The most commonly reported adverse reactions in patients are injection site pain, redness, and a metallic taste in the mouth.

Axumin is marketed by Blue Earth Diagnostics, Ltd., Oxford, United Kingdom

Patent

http://www.google.com/patents/WO2014023775A1?cl=en

The non-natural amino acid [ F]-l-amino-3-fluorocyclobutane-l-carboxylic acid

([¹⁸F]-FACBC, also known as [¹⁸F]-Fluciclovine) is taken up specifically by amino acid transporters and has shown promise for tumour imaging with positron emission tomography (PET).

A known synthesis of [¹⁸F]-FACBC begins with the provision of the protected precursor compound 1 -(N-(t-butoxycarbonyl)amino)-3 –

[((trifluoromethyl)sulfonyl)oxy]-cyclobutane-l-carboxylic acid ethyl ester. This precursor compound is first labelled with [¹⁸F]-fluoride:

II before removal of the two protecting groups:

IT III

EP2017258 (Al) teaches removal of the ethyl protecting group by trapping the [¹⁸F]- labelled precursor compound (II) onto a solid phase extraction (SPE) cartridge and incubating with 0.8 mL of a 4 mol/L solution of sodium hydroxide (NaOH). After 3 minutes incubation the NaOH solution was collected in a vial and a further 0.8 mL 4 mol/L NaOH added to the SPE cartridge to repeat the procedure. Thereafter the SPE cartridge was washed with 3 mL water and the wash solution combined with the collected NaOH solution. Then 2.2 mL of 6 mol/L HCl was then added with heating to 60°C for 5 minutes to remove the Boc protecting group. The resulting solution was purified by passing through (i) an ion retardation column to remove Na⁺ from excess NaOH and Cl^~ from extra HCl needed to neutralise excess of NaOH to get a highly acidic solution before the acidic hydrolysis step, (ii) an alumina column, and (iii) a reverse-phase column. There is scope for the deprotection step(s) and/or the

purification step in the production of [¹⁸F]-FACBC to be simplified.

Example 1: Synthesis of f FIFACBC

No-carrier- added [¹⁸F]fluoride was produced via the ¹⁸0(p,n)¹⁸F nuclear reaction on a GE PETtrace 6 cyclotron (Norwegian Cyclotron Centre, Oslo). Irradiations were performed using a dual-beam, 30μΑ current on two equal Ag targets with HAVAR foils using 16.5 MeV protons. Each target contained 1.6 ml of > 96% [¹⁸0]water (Marshall Isotopes). Subsequent to irradiation and delivery to a hotcell, each target was washed with 1.6 ml of [¹⁶0]water (Merck, water for GR analysis), giving approximately 2-5 Gbq in 3.2 ml of [¹⁶0]water. All radiochemistry was performed on a commercially available GE FASTlab™ with single-use cassettes. Each cassette is built around a one-piece-moulded manifold with 25 three-way stopcocks, all made of polypropylene. Briefly, the cassette includes a 5 ml reactor (cyclic olefin copolymer), one 1 ml syringe and two 5 ml syringes, spikes for connection with five prefilled vials, one water bag (100 ml) as well as various SPE cartridges and filters. Fluid paths are controlled with nitrogen purging, vacuum and the three syringes. The fully automated system is designed for single-step fluorinations with cyclotron-produced [¹⁸F]fluoride. The FASTlab was programmed by the software package in a step-by-step time-dependent sequence of events such as moving the syringes, nitrogen purging, vacuum, and temperature regulation. Synthesis of

[¹⁸F]FACBC followed the three general steps: (a) [¹⁸F]fluorination, (b) hydrolysis of protection groups and (c) SPE purification.

Vial A contained K₂₂₂ (58.8 mg, 156 μπιοΐ), K₂C0₃ (8.1 mg, 60.8 μπιοΐ) in 79.5% (v/v)

MeCN(a_q) (1105 μΐ). Vial B contained 4M HC1 (2.0 ml). Vial C contained MeCN

(4.1ml). Vial D contained the precursor (48.4 mg, 123.5 μιηοΐ) in its dry form (stored at -20 °C until cassette assembly). Vial E contained 2 M NaOH (4.1 ml). The 30 ml product collection glass vial was filled with 200 mM trisodium citrate (10 ml). Aqueous

[¹⁸F]fluoride (1-1.5 ml, 100-200 Mbq) was passed through the QMA and into the ¹⁸0-

H₂0 recovery vial. The QMA was then flushed with MeCN and sent to waste. The trapped [¹⁸F]fluoride was eluted into the reactor using eluent from vial A (730 μΐ) and then concentrated to dryness by azeotropic distillation with acetonitrile (80 μΐ, vial C). Approximately 1.7 ml of MeCN was mixed with precursor in vial D from which 1.0 ml of the dissolved precursor (corresponds to 28.5 mg, 72.7 mmol precursor) was added to the reactor and heated for 3 min at 85°C. The reaction mixture was diluted with water and sent through the tC18 cartridge. Reactor was washed with water and sent through the tC18 cartridge. The labelled intermediate, fixed on the tC18 cartridge was washed with water, and then incubated with 2M NaOH (2.0 ml) for 5 min after which the 2M NaOH was sent to waste. The labelled intermediate (without the ester group) was then eluted off the tC18 cartridge into the reactor using water. The BOC group was hydrolysed by adding 4M HC1 (1.4 ml) and heating the reactor for 5 min at 60 °C. The reactor content with the crude [¹⁸F]FACBC was sent through the HLB and Alumina cartridges and into the 30 ml product vial. The HLB and Alumina cartridges were washed with water (9.1 ml total) and collected in the product vial. Finally, 2M NaOH (0.9 ml) and water (2.1 ml) was added to the product vial, giving a purified formulation of [¹⁸F]FACBC with a total volume of 26 ml. Radiochemical purity was measured by radio-TLC using a mixture of MeCN:MeOH:H₂0:CH₃COOH (20:5:5: 1) as the mobile phase. The radiochemical yield (RCY) was expressed as the amount of radioactivity in the [¹⁸F]FACBC fraction divided by the total used [¹⁸F]fluoride activity (decay corrected). Total synthesis time was 43 min.

The RCY of [¹⁸F]FACBC was 62.5% ± 1.93 (SD), n=4.

/////FDA, diagnostic imaging agent, recurrent prostate cancer, fda 2016, Axumin, marketed, Blue Earth Diagnostics, Ltd., Oxford, United Kingdom, fluciclovine F 18

C1[C@@](C[C@H]1[18F])(N)C(=O)O

Flow synthesis of Fluoxetine

Uncategorized Comments Off

May 272016

Scheme 1: Flow synthesis of fluoxetine (46).

PIC CREDIT, The synthesis of active pharmaceutical ingredients (APIs) using continuous flow chemistry, Marcus Baumann and Ian R. Baxendale, Beilstein J. Org. Chem. 2015, 11, 1194–1219.,doi:10.3762/bjoc.11.134

One of the early published examples of industry-based research on multi-step flow synthesis of a pharmaceutical was reported in 2011 by scientists from Eli Lilly/UK and detailed the synthesis of fluoxetine 46, the API of Prozac[1]. In this account each step was performed and optimised individually in flow, with analysis and purification being accomplished off-line. The synthesis commences with the reduction of the advanced intermediate ketone 47 using a solution of pre-chilled borane–THF complex (48) to yield alcohol 49 (Scheme 1).

Conversion of the pendant chloride into iodide 51 was attempted via Finckelstein conditions, however, even when utilising phase-transfer conditions in order to maintain a homogeneous flow regime the outcome was not satisfactory giving only low conversions. Alternatively direct amination of chloride 49 utilising high temperature flow conditions (140 °C) allowed the direct preparation of amine 50 in excellent yield.

Flow processing using a short residence time (10 min) at the elevated temperature allowed for a good throughput; in addition, the handling of the volatile methylamine within the confines of the flow reactor simplifies the practical aspects of the transformation, however, extra precautions were required in order to address and remove any leftover methylamine that would pose a significant hazard during scaling up.

The final arylation of 50 was intended to be performed as a S_NAr reaction, however, insufficient deprotonation of the alcohol 50 under flow conditions (NaHMDS or BEMP instead of using a suspension of NaH as used in batch) required a modification to the planned approach. To this end a Mitsunobu protocol based on the orchestrated mixing of four reagent streams (50, 54 and reagents 52 and 53) was developed and successfully applied to deliver fluoxetine (46) in high yield.

Overall, this study is a good example detailing the intricacies faced when translating an initial batch synthesis into a sequence of flow steps for which several adaptations regarding choice of reagents and reaction conditions are mandatory in order to succeed.

Ahmed-Omer, B.; Sanderson, A. J. Org. Biomol. Chem. 2011, 9, 3854–3862. doi:10.1039/C0OB00906G
Paper

Preparation of fluoxetine by multiple flow processing steps

Batoul Ahmed-Omer^a and Adam J. Sanderson*^a

*Corresponding authors

^aEli Lilly and Co. Ltd., Lilly Research Centre, Erl Wood Manor, Windlesham, Surrey, UK

Org. Biomol. Chem., 2011,9, 3854-3862

DOI: 10.1039/C0OB00906G

http://pubs.rsc.org/en/Content/ArticleLanding/2011/OB/c0ob00906g#!divAbstract

Microflow technology is established as a modern and fashionable tool in synthetic organic chemistry, bringing great improvement and potential, on account of a series of advantages over flask methods. The study presented here focuses on the application of flow chemistry process in performing an efficient multiple step syntheses of (±)-fluoxetine as an alternative to conventional synthetic methods, and one of the few examples of total synthesis accomplished by flow technique.

Graphical abstract: Preparation of fluoxetine by multiple flow processing steps

1 The general method set-up of flow process used for the synthesis of (±)- fluoxetine.

Scheme 1 Synthesis of (±)-fluoxetine in flow: (i) BH3·THF, r.t., 5 min (77%); (ii) NaI, toluene: water, 100 °C, 20 min (43%); (iii); MeNH2 (aq), …

//////////Flow synthesis, fluoxetine

SETIPIPRANT

Uncategorized Comments Off

May 272016

Setipiprant, KYTH-105

CAS 866460-33-5

2-(2-(1-naphthoyl)-8-fluoro-1,2,3,4-tetrahydropyrido[4,3-b]indol-5-yl)acetic acid

2-[8-fluoro-2-(naphthalene-1-carbonyl)-3,4-dihydro-1H-pyrido[4,3-b]indol-5-yl]acetic acid

5H-Pyrido(4,3-b)indole-5-acetic acid, 8-fluoro-1,2,3,4-tetrahydro-2-(1-naphthalenylcarbonyl)-

MF C24H19FN2O3

MW 402.4176632

IND FILED BY ALLERGAN FOR Alopecia

ACT-129968, a CRTH2 receptor antagonist, had been in phase II clinical trials at Actelion

Setipiprant; UNII-BHF20LA2GM; ACT-129968; 866460-33-5;

Setipiprant is a prostaglandin D2 (PGD2) antagonist. Essentially, it inhibits PGD2 receptor activity

KYTH-105 had previously been studied as a potential allergic inflammation treatment and had undergone eight clinical trials, resulting in a safety database of more than 1,000 patients. Treatment in all studies was well tolerated across all treatment groups.

Intellectual Property
KYTHERA acquired exclusive worldwide rights to KYTH-105, as well as certain patent rights covering the use of PGD2 receptor antagonists for the treatment of hair loss (often presenting as male pattern baldness, or androgenic alopecia).

Next Steps
KYTHERA plans to file an Investigational New Drug (IND) application and initiate a proof-of-concept study to establish the efficacy of KYTH-105 in male subjects with androgenic alopecia (AGA).

In 2015, Allergan acquired Kythera.

2-(2-(1-Naphthoyl)-8-fluoro-3,4-dihydro-1H-pyrido[4,3-b]indol-5(2H)-yl)acetic Acid

mp 224.0 °C.

LC(1)/ESI-MS t_R = 0.83 min; m/z [M + H⁺] = 403.09.

¹H NMR (DMSO-d₆), 65:35 mixture of two rotamers, δ: 8.02 (m, 2 H), 7.76 (d, J = 7.8 Hz, 0.65 H), 7.72 (m, 0.35 H), 7.49–7.64 (m, 3.35 H), 7.35–7.49 (m, 2.35 H), 6.98 (ddd, J_H–F = 9.3 Hz, J₁ = 9.3 Hz, J₂ = 2.4 Hz, 0.65 H), 6.88 (m, 0.65 H), 4.85–5.14 (m, 3.3 H), 4.42 (m, 0.35 H), 4.32 (m, 0.7 H), 4.06 (m, 0.35 H), 3.50 (t, J = 5.5 Hz, 1.3 H), 2.95 (m, 0.70 H), 2.68 (m, 0.65 H), 2.58 (m, 0.65 H).

¹³C NMR (DMSO-d₆) δ: 170.7, 169.2, 157.7 (d, J_C–F = 232 Hz), 157.4 (d, J_C–F = 233 Hz), 137.1, 136.2, 135.1, 134.9, 134.0, 133.8, 133.5, 129.6, 129.5, 129.4, 129.3, 128.9, 128.8, 127.5, 127.4, 127.0, 126.9, 126.0, 125.9, 125.7 (d, J_C–F = 10 Hz), 125.2, 125.1, 125.0, 124.1, 123.9, 110.9 (d, J_C–F = 10 Hz), 110.8 (m), 109.3 (d, J_C–F = 26 Hz), 109.1 (d, J_C–F = 26 Hz), 106.7 (m), 103.3 (d, J_C–F = 23 Hz), 103.0 (d, J_C–F = 23 Hz), 44.73, 44.70, 44.5, 44.4, 39.5, 39.3, 23.1, 22.3.

HRMS (ESI): m/zcalcd for C₂₄H₂₀N₂O₃F [M + H⁺] 403.1458, found 403.1458.

SYNTHERSIS

Setipiprant (INN) (developmental code names ACT-129,968, KYTH-105) is a drug originally developed by Actelion which acts as a selective, orally available antagonist of the prostaglandin D₂ receptor 2 (DP₂).^[1] It was initially researched as a treatment for allergies and inflammatory disorders, particularly asthma, but despite being well tolerated in clinical trials and showing reasonable efficacy against allergen-induced airway responses in asthmatic patients,^[2]^[3] it failed to show sufficient advantages over existing drugs and was discontinued from further development in this application.^[4]

However, following the discovery in 2012 that the prostaglandin D₂ receptor (DP/PGD₂) is expressed at high levels in the scalp of men affected by male pattern baldness,^[5] the rights to setipiprant were acquired by Kythera with a view to potentially developing this drug as a novel treatment for baldness, with a previously unexploited mechanism of action.^[6] While it is too early to tell whether setipiprant will be an effective treatment for this condition, the favorable pharmacokinetics and relative lack of side effects seen in earlier clinical trials mean that fresh clinical trials for this new application can be conducted fairly quickly.^[7]

Prostaglandin D2 is a known agonist of the thromboxane A2 (TxA2) receptor, the PGD2 (DP) receptor and the recently identified G-protein-coupled “chemoattractant receptor- homologous molecule expressed on Th2 cells” (CRTH2).

The response to allergen exposure in a previously sensitized host results in a cascade effect involving numerous cell types and release of a number of cytokines, chemokines, and multiple mediators. Among these critical initiators are the cytokines interleukin (IL)-4, IL-13, and IL-5, which play critical roles in Th2 cell differentiation, immunoglobulin (Ig)E synthesis, mast cell growth and differentiation, upregulation of CD23 expression, and the differentiation, recruitment, and activation of eosinophils. The stimulated release of the array of mediators, causes end-organ damage, including constriction and hyperresponsi- veness, vascular permeability, edema, mucous secretion, and further inflammation.

Because of the number of responses targeted, corticosteroids have proven to be the most effective therapy. Rather than antagonizing these specific responses in a directed way, another approach is to alter the immune response, that is, to change the nature of the immunological response to allergen. CRTH2 is preferentially expressed on Th2 cells and is a chemoattractant receptor for PGD2 that mediates PGD2-dependent migration of blood Th2 cells. Chemoattractants are responsible for the recruitment of both Th2 cells and other effector cells of allergic inflammation, which can provide the conceptual basis for the development of new therapeutic strategies in allergic conditions.

So far, few compounds having CRTH2 antagonistic activity have been reported in the patent literature. Bayer AG claims the use of Ramatroban ((3R)-3-(4-fluorobenzene- sulfonamido)-l,2,3,4-tetrahydrocarbazole-9-propionic acid) for the prophylaxis and treatment of allergic diseases, such as asthma, allergic rhinitis or allergic conjuvatitis

(GB 2388540). Further, (2-tert.-butoxycarbonyl-l, 2, 3, 4-tetrahydro-pyrido[4,3-b]indol-5- yl)-acetic acid and (2-ethoxycarbonyl-l, 2, 3, 4-tetrahydro-pyrido[4,3-b]indol~5-yl)-acetic acid are disclosed by Kyle F. et al in two patent applications (US 5817756 and WO 9507294, respectively).

Furthermore, oral bioavailability of the Ramatroban and its ability to inhibit prostaglandin D2-induced eosinophil migration in vitro has been reported (Journal of Pharmacology and Experimental Therapeutics, 305(1), p.347-352 (2003)).

Description of the invention:

It has now been found that compounds of the general Formulae (I) and (II) of the present invention are CRTH2 receptor antagonists. These compounds are useful for the treatment of both chronic and acute allergic/immune disorders such as allergic asthma, rhinitis, chronic obstructive pulmonary disease (COPD), dermatitis, inflammatory bowel disease, rheumatoid arthritis, allergic nephritis, conjunctivitis, atopic dermatitis, bronchial asthma, food allergy, systemic mast cell disorders, anaphylactic shock, urticaria, eczema, itching, inflammation, ischemia-reperfusion injury, cerebrovascular disorders, pleuritis, ulcerative colitis, eosinophil-related diseases, such as Churg-Strauss syndrome and sinusitis, basophil- related diseases, such as basophilic leukemia and basophilic leukocytosis.

The compounds of general Formulae (I) and (II), especially those mentioned as being preferred, display high selectivity towards the CRTH2 receptor. No antagonistic effects (IC₅₀ >10 μM) are observed on e.g. prostaglandin D2 receptor DPI; PGI2 receptor (IP), PGE2 receptors (EPl, EP2, EP3, EP4), PGF2 receptor (FP), thromboxane receptor A2 (TxA2), leukotriene receptors (CysLTl, CysLT2, LTB4), complement receptor (C5a), angiotensin receptors (ATI, AT2) or serotonin receptor 5HT2c.

The solubility of compounds of general Formulae (I) and (II) in buffer at pH 7 is generally >800 μg/ml.

In vitro assays with rat and dog liver microsomes, or with rat and human hepatocytes revealed high metabolic stability for compounds of general Foπnulae (I) and (II), especially for those compounds mentioned as being preferred.

The compounds of general Formulae (I) and (II), especially those mentioned as being preferred, do not interfere with cytochrome P-450 enzymes, e.g. they are neither degraded by, nor do they inhibit such enzymes.

Excellent pharmacokinetic profiles have been observed for compounds of general Formulae (I) and (II), especially for those compounds mentioned as being preferred, after oral administration (10 mg/kg) to rats and dogs (bioavailability 20-80%, T_max 30 min, C_max 2000- 6000 ng/ml, low clearance, T_{] 2}4-8 h). The compounds of general Formulae (I) and (II), especially those mentioned as being preferred, are efficacious in vitro, inhibiting PGD2-induced migration of eosinophils or other CRTH2 expressing cells in a cell migration assay. A number of techniques have been developed to assay such chemotactic migration (see, e.g., Leonard et al., 1995, “Measurement of α- and β-Chemokines”, in Current Protocols in Immunology, 6.12.1- 6.12.28, Ed. Coligan et al, John Wiley & Sons, Inc. 1995). The compounds of the present invention are tested using a protocol according to H. Sugimoto et al. (J Pharmacol Exp Ther. 2003, 305(1), 347-52), or as described hereinafter: Purified eosinophils are labeled with a fluorescent dye, i.e. Calcein-AM and loaded in BD Falcon FluoroBlock upper inserts. Test compounds are diluted and incubated with eosinophils in the BD Falcon

FluoroBlock upper inserts for 30 min at 37 °C in a humidified CO₂ incubator. A constant amount of PGD2 is added to BD Falcon FluoroBlock lower chamber, at a concentration known to have a chemotactic effect on CRTH2 cells. As a control, at least one aliquot in the upper well does not contain test compound. The inserts are combined with the chambers and are incubated for 30 min at 37 °C in a humidified CO₂ incubator. After an incubation period, the number of migrating cells on the lower chamber is counted using a fluorescent reader, i.e. an Applied Biosystems Cyto Fluor 4000 plate reader. The contribution of a test compound to the chemotactic activity of PGD2 is measured by comparing the chemotactic activity of the aliquots containing only dilution buffer with the activity of aliquots containing a test compound. If addition of the test compound to the solution results in a decrease in the number of cells detected in the lower chamber relative to the number of cells detected using a solution containing only PGD2, then there is identified an antagonist of PGD2 induction of chemotactic activity of eosinophils.

PAPER

Journal of Medicinal Chemistry (2013), 56(12), 4899-4911

http://pubs.acs.org/doi/abs/10.1021/jm400122f

Identification of 2-(2-(1-Naphthoyl)-8-fluoro-3,4-dihydro-1H-pyrido[4,3-b]indol-5(2H)-yl)acetic Acid (Setipiprant/ACT-129968), a Potent, Selective, and Orally Bioavailable Chemoattractant Receptor-Homologous Molecule Expressed on Th2 Cells (CRTH2) Antagonist

Heinz Fretz*, Anja Valdenaire, Julien Pothier, Kurt Hilpert, Carmela Gnerre, Oliver Peter, Xavier Leroy, and Markus A. Riederer

Drug Discovery Unit, Actelion Pharmaceuticals Ltd., Gewerbestrasse 16, CH-4123 Allschwil, Switzerland

J. Med. Chem., 2013, 56 (12), pp 4899–4911

DOI: 10.1021/jm400122f

Herein we describe the discovery of the novel CRTh2 antagonist 2-(2-(1-naphthoyl)-8-fluoro-3,4-dihydro-1H-pyrido[4,3-b]indol-5(2H)-yl)acetic acid 28 (setipiprant/ACT-129968), a clinical development candidate for the treatment of asthma and seasonal allergic rhinitis. A lead optimization program was started based on the discovery of the recently disclosed CRTh2 antagonist 2-(2-benzoyl-3,4-dihydro-1H-pyrido[4,3-b]indol-5(2H)-yl)acetic acid 5. An already favorable and druglike profile could be assessed for lead compound 5. Therefore, the lead optimization program mainly focused on the improvement in potency and oral bioavailability. Data of newly synthesized analogs were collected from in vitro pharmacological, physicochemical, in vitro ADME, and in vivo pharmacokinetic studies in the rat and the dog. The data were then analyzed using a traffic light selection tool as a visualization device in order to evaluate and prioritize candidates displaying a balanced overall profile. This data-driven process and the excellent results of the PK study in the rat (F = 44%) and the dog (F = 55%) facilitated the identification of 28 as a potent (IC₅₀ = 6 nM), selective, and orally available CRTh2 antagonist.

PAtent

WO 2005095397

http://www.google.co.in/patents/WO2005095397A1?cl=en

Formula 6.

Scheme 1

Step a)

Step b)

Scheme 2

Formula (I).

References

Fretz H, Valdenaire A, Pothier J, Hilpert K, Gnerre C, Peter O, Leroy X, Riederer MA. Identification of 2-(2-(1-naphthoyl)-8-fluoro-3,4-dihydro-1H-pyrido[4,3-b]indol-5(2H)-yl)acetic acid (setipiprant/ACT-129968), a potent, selective, and orally bioavailable chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) antagonist. J Med Chem. 2013 Jun 27;56(12):4899-911. doi: 10.1021/jm400122f PMID 23721423
Sidharta PN, Diamant Z, Dingemanse J. Single- and multiple-dose tolerability and pharmacokinetics of the CRTH2 antagonist setipiprant in healthy male subjects. Fundam Clin Pharmacol. 2014 Dec;28(6):690-9. doi: 10.1111/fcp.12079 PMID 24734908
Diamant Z, Sidharta PN, Singh D, O’Connor BJ, Zuiker R, Leaker BR, Silkey M, Dingemanse J. Setipiprant, a selective CRTH2 antagonist, reduces allergen-induced airway responses in allergic asthmatics. Clin Exp Allergy. 2014 Aug;44(8):1044-52. doi: 10.1111/cea.12357 PMID 24964348
Norman P. Update on the status of DP2 receptor antagonists; from proof of concept through clinical failures to promising new drugs. Expert Opin Investig Drugs. 2014 Jan;23(1):55-66. doi: 10.1517/13543784.2013.839658 PMID 24073896
Garza LA, et al. Prostaglandin D2 inhibits hair growth and is elevated in bald scalp of men with androgenetic alopecia. Science Translational Medicine, 21 March 2012; 4(126):126ra34. doi: 10.1126/scitranslmed.3003122
George Cotsarelis, Garret Fitzgerald, Luis Garza. Compositions and methods for regulating hair growth. US Patent application 2015/0072963
Pipeline KYTH-105 (setipiprant)
http://files.shareholder.com/downloads/AMDA-MFNLA/4023632629x0x817836/4E5AC47A-B9EE-4296-9D97-631C0F6B7C97/KYTH-105_setipiprant_.pdf

Patent ID	Date	Patent Title
US2015072963	2015-03-12	COMPOSITIONS AND METHODS FOR REGULATING HAIR GROWTH
US2014328861	2014-11-06	Combination of CRTH2 Antagonist and a Proton Pump Inhibitor for the Treatment of Eosinophilic Esophagitis
US2010234396	2010-09-16	Tetrhydropyridoindole Derivatives
US7714132	2010-05-11	Tetrahydropyridoindole derivatives

S etipiprant
Systematic (IUPAC) name

2-[8-fluoro-2-(naphthalene-1-carbonyl)-3,4-dihydro-1H-pyrido[4,3-b]indol-5-yl]acetic acid
Clinical data
Administration	Oral
Identifiers
CASRN	866460-33-5
ATC code	none
PubChem	CID 49843471
Chemical data
Formula	C₂₄H₁₉FN₂O₃
Molar mass	402.417 g/mol

///////Setipiprant, KYTH-105, 866460-33-5, ALLERGAN, Alopecia, KYTHERA

c15ccccc5cccc1C(=O)N(CC3)Cc2c3n(CC(O)=O)c(cc4)c2cc4F

CFI-402257

Uncategorized Comments Off

May 252016

CFI-402257

N-cyclopropyl-4-(7-((((1s,3s)-3-hydroxy-3-methylcyclobutyl)methyl)amino)-5-(pyridin-2-yloxy)pyrazolo[1,5-a]pyridin-3-yl)-2-methylbenzamide

N-cyclopropyl-4-(7-( (((Is, 3s)-3-hydroxy-3-methylcyclobutyl)methyl)amino)-5- (pyridin-3-yloxy)pyrazolol 1 , 5-a ]pyrimidin-3-yl)-2-methylbenzamide

CAS 1610759-22-2 (free base); 1610677-37-6 (HCl)
MF: C29H31N5O3
MW: 497.2427

University Health Network

CFI-402257 is a highly potent and selective TTK (threonine tyrosine kinase) Inhibitor ((TTK Ki = 0.1 nM) with potential anticancer activity. TTK is an essential chromosomal regulator and is overexpressed in aneuploid tumors. High TTK levels correlate with a high tumor grade11 and poor patient outcomes. TTK inhibition are associated with a disabled mitotic checkpoint, resulting in chromosome segregation errors, aneuploidy, and cell death.

Synthesis

SYN OF INTERMEDIATE

SYNTHESIS COLOUR INDICATED

SYN OF INTERMEDIATES

IF YOU HAVE ENJOYED IT ………EMAIL ME amcrasto@gmail.com, +919323115463, India

INDIA FLAG

DR ANTHONY CRASTO , WORLDDRUGTRACKER, HELPING MILLIONS, MAKING INDIA AND INDIANS PROUD

Protein kinases have been the subject of extensive study in the search for new therapeutic agents in various diseases, for example, cancer. Protein kinases are known to mediate intracellular signal transduction by effecting a phosphoryl transfer from a nucleoside triphosphate to a protein acceptor that is involved in a signaling pathway. There are a number of kinases and pathways through which extracellular and other stimuli cause a variety of cellular responses to occur inside the cell.

Human TTK protein kinase (TTK), also known as tyrosine threonine kinase, dual specificity protein kinase TTK, Monopolar Spindle 1 (Mpsl) and Phosphotyrosine -Picked Threonine Kinase (PYT), is a conserved multispecific kinase that is capable of phosphorylating serine, threonine and tyrosine residues when expressed in E. coli (Mills et al., J. Biol. Chem. 22(5): 16000-16006 (1992)). TTK mRNA is not expressed in the majority of physiologically normal tissues in human (Id). TTK mRNA is expressed in some rapidly proliferating tissues, such as testis and thymus, as well as in some tumors (for example, TTK mRNA was not expressed in renal cell carcinoma, was expressed in 50% of breast cancer samples, was expressed in testicular tumors and ovarian cancer samples) (Id). TTK is expressed in some cancer cell lines and tumors relative to normal counterparts (Id.; see also WO 02/068444 Al).

Therefore, agents which inhibit a protein kinase, in particular TTK, have the potential to treat cancer. There is a need for additional agents which can act as protein kinase inhibitors, in particular TTK inhibitors.

In addition, cancer recurrence, drug resistance or metastasis is one of the major challenges in cancer therapies. Cancer patients who responded favorably to the initial anticancer therapy often develop drug resistance and secondary tumors that lead to the relapse of the disease. Recent research evidences suggest that the capability of a tumor to grow and propagate is dependent on a small subset of cells within the tumor. These cells are termed tumor-initiating cells (TICs) or cancer stem cells. It is thought that the TICs are responsible for drug resistance, cancer relapse and metastasis. Compounds that can inhibit the growth and survival of these tumor-initiating cells can be used to treat cancer, metastasis or prevent recurrence of cancer. Therefore, a need exists for new compounds that can inhibit the growth and survival of tumor- imitating cells.

PATENT

WO 2015070349

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2015070349&recNum=1&maxRec=&office=&prevFilter=&sortOption=&queryString=&tab=PCTDescription

A4: N-cyclopropyl-4-(7-( (((Is, 3s)-3-hydroxy-3-methylcyclobutyl)methyl)amino)-5- (pyridin-3-yloxy)pyrazolol 1 , 5-a ]pyrimidin-3-yl)-2-methylbenzamide hydrochloride and its free base

A). Through Boc deprotection: A mixture of tert-butyl (3- bromo-5-(pyridin-3-yloxy)pyrazolo[l,5-a]pyrimidin-7- yl)(((ls,3s)-3-((tert-butoxycarbonyl)oxy)-3- methylcyclobutyl)methyl)carbamate (0.23 g, 0.38 mmol), N- cyclopropyl-2-methyl-4-(4,4,5,5-tetramethyl-l,3,2-

dioxaborolan-2-yl)benzamide (0.15 g, 0.49 mmol), PdC dppfDCM (0.15 g, 0.49 mmol), and 2M K₃P0₄ (0.57 mL, 1.14 mmol) in THF (4 mL) was charged with Ar and heated in the microwave at 130 °C for 3 h. Water and EtOAc were added to separate the phases and the aqueous phase was extracted with EtOAc. The combined organic extracts were dried over NaSC>4, filtered and concentrated. The crude product was purified by flash chromatography (gradient: EtOAc/hex 20-60%) to give a yellow oil.

The above intermediate was dissolved in DCM (10 mL) and treated with TFA (3 mL) at rt for 3 h. After reaction completion, solvent was removed in vacuo and the crude product was dissolved in MeOH (5 mL). The mixture was filtered and purified by prep-HPLC. The compound was passed through a PoraPak cartridge and triturated with Et₂0 to give the title compound as a free base (white solid). The free base was dissolved in MeOH (5 mL), and HC1 (1 M Et₂0, 2 equiv) was then added slowly. Solvent was removed in vacuo to give the title compound as a beige solid in HC1 salt (96 mg, 47% over 2 steps). ¾ NMR (400 MHz, CD₃OD) δ ppm 9.14 (br. s, 1H), 8.89-8.82 (m, 1H), 8.79-8.71 (m, 1H), 8.40 (s, 1H), 8.31-8.21 (m, 1H), 7.68 (s, 1H), 7.59 (d, J = 9.5 Hz, 1H), 7.23 (d, J= 8.0 Hz, 1H), 6.06 (s, 1H), 3.56 (d, J= 6.5 Hz, 2H), 2.88-2.79 (m, 1H),

2.40-2.31 (m, 1H), 2.29 (s, 3H), 2.26-2.18 (m, 2H), 1.99-1.89 (m, 2H), 1.37 (s, 3H),

0.85-0.76 (m, 2H), 0.63-0.53 (m, 2H); MS ESI [M + H]⁺ 499.3, calcd for [C^HsoNeOs +

H]⁺ 499.2. HPLC purity: 99.5% at 254 nm.

B). Through PMB deprotection: A mixture of N- cyclopropyl-4-(7-((((ls,3s)-3-hydroxy-3- methylcyclobutyl)methyl)(4-methoxybenzyl)amino)-5- (pyridin-3-yloxy)pyrazolo[l,5-a]pyrimidin-3-yl)-2- methylbenzamide (9.6 g, 15.5 mmol), TFA (50 mL) in DCE

(70 mL) was heated in an oil bath at 50 °C for 4 h. After reaction completion, solvent was removed in vacuo and the crude product was dissolved in a mixture of MeOH/DCM (100 mL/25 mL). 2M Na₂CC (150 mL) was then added and the resulting mixture was stirred at rt for 30 min. The reaction mixture was diluted with DCM and the phases were separated. The aqueous phase was extracted with DCM and the combined organic extracts were washed with water, dried over MgSC , filtered and concentrated. The crude product was triturated and sonicated in a mixture of DCM/Et₂0 (10 mL/70 mL) to give the title compound as a off white solid in free base (5.9 g, 77%). Ti NMR (400 MHz, CD₃OD) δ ppm 8.58-8.53 (m, 1H), 8.50-8.46 (m, 1H), 8.36 (s, 1H), 7.86-7.80 (m, 1H), 7.76-7.72 (m, 1H), 7.61-7.55 (m, 2H), 7.18 (d, J = 8.0 Hz, 1H), 5.92 (s, 1H), 3.52 (d, J = 6.8 Hz, 2H), 2.86-2.77 (m, 1H), 2.38-2.28 (m, 1H), 2.25 (s, 3H), 2.24-2.18 (m, 2H), 1.99-1.88 (m, 2H), 1.37 (s, 3H), 0.84-0.75 (m, 2H), 0.64-0.54 (m, 2H); MS ESI [M + H]⁺ 499.2, calcd for [CzsHsoNgOs + H]⁺ 499.2. HPLC purity: 96.1% at 235 nm.

PATENT

WO 2014075168

PAPER

http://pubs.acs.org/doi/abs/10.1021/acsmedchemlett.5b00485

This work describes a scaffold hopping exercise that begins with known imidazo[1,2-a]pyrazines, briefly explores pyrazolo[1,5-a][1,3,5]triazines, and ultimately yields pyrazolo[1,5-a]pyrimidines as a novel class of potent TTK inhibitors. An X-ray structure of a representative compound is consistent with 1¹/₂ type inhibition and provides structural insight to aid subsequent optimization of in vitro activity and physicochemical and pharmacokinetic properties. Incorporation of polar moieties in the hydrophobic and solvent accessible regions modulates physicochemical properties while maintaining potency. Compounds with enhanced oral exposure were identified for xenograft studies. The work culminates in the identification of a potent (TTK K_i = 0.1 nM), highly selective, orally bioavailable anticancer agent (CFI-402257) for IND enabling studies.

Discovery of Pyrazolo[1,5-a]pyrimidine TTK Inhibitors: CFI-402257 is a Potent, Selective, Bioavailable Anticancer Agent

Yong Liu^†, Radoslaw Laufer^†, Narendra Kumar Patel^†, Grace Ng^†, Peter B. Sampson^†, Sze-Wan Li^†, Yunhui Lang^†,Miklos Feher^†, Richard Brokx^†, Irina Beletskaya^†, Richard Hodgson^†, Olga Plotnikova^†, Donald E. Awrey^†, Wei Qiu^‡, Nickolay Y. Chirgadze^‡, Jacqueline M. Mason^†, Xin Wei^†, Dan Chi-Chia Lin^†, Yi Che^†, Reza Kiarash^†, Graham C. Fletcher^†, Tak W. Mak^†, Mark R. Bray^†, and Henry W. Pauls^*^†

^† Campbell Family Institute for Breast Cancer Research, University Health Network, TMDT East Tower, MaRS Centre, 101 College Street, Toronto, Ontario M5G 1L7, Canada

^‡ Campbell Family Cancer Research Institute, University Health Network, Princess Margaret Cancer Center, 610 University Avenue, Toronto, Ontario M5G 2C4, Canada

ACS Med. Chem. Lett., Article ASAP

DOI: 10.1021/acsmedchemlett.5b00485

*E-mail: henry.pauls@cogeco.ca. Phone: 905-337-3446.

REFERENCES

Discovery of Pyrazolo[1,5-a]pyrimidine TTK Inhibitors: CFI-402257 is a Potent, Selective, Bioavailable Anticancer Agent
Yong Liu, Radoslaw Laufer, Narendra Kumar Patel, Grace Ng, Peter B. Sampson, Sze-Wan Li, Yunhui Lang, Miklos Feher, Richard Brokx, Irina Beletskaya, Richard Hodgson, Olga Plotnikova, Donald E. Awrey, Wei Qiu, Nickolay Y. Chirgadze, Jacqueline M. Mason, Xin Wei, Dan Chi-Chia Lin, Yi Che, Reza Kiarash, Graham C. Fletcher, Tak W. Mak, Mark R. Bray, and Henry W. Pauls
Publication Date (Web): May 6, 2016 (Letter)
DOI: 10.1021/acsmedchemlett.5b00485

////TTK inhibitors, CFI-402257, pyrazolo[1,5-a]pyrimidines, 1¹/₂ type inhibitors, 1610759-22-2, 1610677-37-6

C[C@]1(O)C[C@H](C1)CNc2cc(nc3c(cnn23)c5ccc(C(=O)NC4CC4)c(C)c5)Oc6cccnc6

Antimycobacterial Agents

PRECLINICAL, Uncategorized Comments Off

May 252016

Styryl Hydrazine Thiazole Hybrids

Will be updated………kindly email amcrasto@gmail.com

DATA

ABOUT Dehydrozingerone

Dehydrozingerone; Feruloylmethane; 1080-12-2; 4-(4-Hydroxy-3-methoxyphenyl)-3-buten-2-one; 4-(4-hydroxy-3-methoxyphenyl)but-3-en-2-one; Vanillalacetone;

http://pubs.acs.org/doi/abs/10.1021/np300465f

J. Nat. Prod., 2012, 75 (12), pp 2088–2093

DOI: 10.1021/np300465f

Dehydrozingerone (1) is a pungent constituent present in the rhizomes of ginger (Zingiber officinale) and belongs structurally to the vanillyl ketone class. It is a representative of half the chemical structure of curcumin (2), which is an antioxidative yellow pigment obtained from the rhizomes of turmeric (Curcuma longa). Numerous studies have suggested that 2 is a promising phytochemical for the inhibition of malignant tumors, including colon cancer. On the other hand, there have been few studies on the potential antineoplastic properties of 1, and its mode of action based on a molecular mechanism is little known. Therefore, the antiproliferative effects of1 were evaluated against HT-29 human colon cancer cells, and it was found that 1 dose-dependently inhibited growth at the G2/M phase with up-regulation of p21. Dehydrozingerone additionally led to the accumulation of intracellular ROS, although most radical scavengers could not clearly repress the cell-cycle arrest at the G2/M phase. Furthermore, two synthetic isomers of1 (iso-dehydrozingerone, 3, and ortho-dehydrozingerone, 4) were also examined. On comparing of their activities, accumulation of intracellular ROS was found to be interrelated with growth-inhibitory effects. These results suggest that analogues of 1 may be potential chemotherapeutic agents for colon cancer

PAPER

Series of styryl hydrazine thiazole hybrids inspired from dehydrozingerone (DZG) scaffold were designed and synthesized by molecular hybridization approach. In vitro antimycobacterial activity of synthesized compounds was evaluated against Mycobacterium tuberculosis H₃₇Rv strain. Among the series, compound 6o exhibited significant activity (MIC = 1.5 μM; IC₅₀ = 0.48 μM) along with bactericidal (MBC = 12 μM) and intracellular antimycobacterial activities (IC₅₀ = <0.098 μM). Furthermore, 6o displayed prominent antimycobacterial activity under hypoxic (MIC = 46 μM) and normal oxygen (MIC = 0.28 μM) conditions along with antimycobacterial efficiency against isoniazid (MIC = 3.2 μM for INH-R1; 1.5 μM for INH-R2) and rifampicin (MIC = 2.2 μM for RIF-R1; 6.3 μM for RIF-R2) resistant strains of Mtb. Presence of electron donating groups on the phenyl ring of thiazole moiety had positive correlation for biological activity, suggesting the importance of molecular hybridization approach for the development of newer DZG clubbed hydrazine thiazole hybrids as potential antimycobacterial agents.

Dehydrozingerone Inspired Styryl Hydrazine Thiazole Hybrids as Promising Class of Antimycobacterial Agents

Girish A. Hampannavar^†, Rajshekhar Karpoormath^*^†, Mahesh B. Palkar^§^†, Mahamadhanif S. Shaikh^†, and Balakumar Chandrasekaran^†

^† Department of Pharmaceutical Chemistry, Discipline of Pharmaceutical Sciences, College of Health Sciences, University of KwaZulu-Natal, Westville Campus, Durban 4000, South Africa

^§ Department of Pharmaceutical Chemistry, K.L.E. University College of Pharmacy, Vidyanagar, Hubballi 580031, Karnataka, India

ACS Med. Chem. Lett., Article ASAP

DOI: 10.1021/acsmedchemlett.6b00088

http://pubs.acs.org/doi/abs/10.1021/acsmedchemlett.6b00088

*Phone: +27 31 260 7179. Fax: +27 (0) 31 260 7792. E-mail: karpoormath@ukzn.ac.za.

///////Antimycobacterial activity, bactericidal, dehydrozingerone, NIAID, thiazole, PRECLINCAL

c1(ccc(c(c1)OC)OC)/C=C/C(C)=N/Nc2nc(cs2)c3ccc(cc3)N

FDA approved a switchover from batch to the new technology for production of HIV drug Prezista, Darunavir on a line at its plant in Gurabo, Puerto Rico

Uncategorized Comments Off

May 232016

Above is an Illustration example,

FDA urges companies to get on board with continuous manufacturing

The FDA gave Johnson & Johnson’s ($JNJ) Janssen drug unit the thumbs up last week for the continuous manufacturing process that it has been working on for 5 years. The agency approved a switchover from batch to the new technology for production of HIV drug Prezista on a line at its plant in Gurabo, Puerto Rico……http://www.fiercepharma.com/manufacturing/fda-urges-companies-to-get-on-board-continuous-manufacturing

Darunavir

SEE……http://www.en-cphi.cn/news/show-29367.html

Just after opening a refurbished manufacturing facility in Cape Town, South Africa earlier this year, pharma giant Johnson & Johnson ($JNJ) recently opened the doors to its Global Public Health Africa Operations office there.

The company has invested $21 million (300 million rand) in the facilities. The global public health facility will focus on HIV, tuberculosis and maternal, newborn and child health, South Africa – The Good News reported.

“This (investment) tells us that South Africa has the capability to provide a facility for world-class manufacturing,” Rob Davies, minister of the Department of Trade and Industry told the publication.

Johnson & Johnson, which has operated in South Africa for more than 86 years, planned to close the Cape Town manufacturing plant by the end of 2008 but was persuaded to keep the facility open for local manufacturing to serve sub-Saharan business. By 2015, the plant was cited by J&J as the most-improved in cost competitiveness from 30 company plants worldwide.

Earlier this month, the FDA gave J&J’s Janssen drug unit the go-ahead to proceed with the continuous manufacturing process it’s been working on for 5 years. The agency approved a switchover from batch to the new technology for production of HIV drug Prezista, Darunavir on a line at its plant in Gurabo, Puerto Rico.

AN EXAMPLE NOT RELATED TO DARUNAVIR

References

May 20-21, 2014 (Link to 2016 Meeting Website)

Continuous Bioprocessing

https://iscmp.mit.edu/white-papers/white-paper-4

READ

Achieving Continuous Manufacturing: Technologies and Approaches for Synthesis, Work-Up and Isolation of Drug Substance

https://iscmp.mit.edu/white-papers/white-paper-1

//////

Higenamine Hydrochloride

Uncategorized Comments Off

May 232016

Higenamine Hydrochloride

6,7-Isoquinolinediol, 1,2,3,4-tetrahydro-1-[(4-hydroxyphenyl)methyl]-, hydrochloride (9CI)
6,7-Isoquinolinediol, 1,2,3,4-tetrahydro-1-[(4-hydroxyphenyl)methyl]-, hydrochloride, (±)-
(±)-Demethylcoclaurine hydrochloride

A β-adrenoceptor partial agonist potentially for the treatment of coronary heart disease.

CAS No.11041-94-4 (Higenamine hydrochloride)

CAS 5843-65-2(free)

Higenamine (norcoclaurine) is a chemical compound found in a variety of plants including Nandina domestica (fruit), Aconitum carmichaelii (root), Asarum heterotropioides, Galium divaricatum (stem and vine), Annona squamosa, and Nelumbo nucifera (lotus seeds).

Legality

Higenamine, also known as norcoclaurine HCl, is legal to use within food supplements in the UK, EU, the USA and Canada. but banned use in The NCAA. Its main is within food supplements developed for weight management, also known as ‘fat burners’. However, products containing (or claiming to contain) pharmacological relevant quantities still require registration as a medicine. The regulatory boundaries for higenamine are unclear as modern formulations have not been clinically evaluated. Traditional formulations with higenamine have been used for thousands of years within Chinese medicine and come from a variety of sources including fruit and orchids. There are no studies comparing the safety of modern formulations (based on synthetic higenamine) with traditional formulations. Nevertheless, it will not be added to the EU ‘novel foods’ catalogue, which details all food supplements that require a safety assessment certificate before use.^[1]

Pharmacology

Since higenamine is present in plants which have a history of use in traditional medicine, the pharmacology of this compound has attracted scientific interest. A variety of effects have been observed in in vitro studies and in animal models, but its effects in humans are unknown.

The results of a 2009 study exposed the compound as a β₂ adrenergic receptor agonist.^[2]

In animal models, higenamine has been demonstrated to be a β₂ adrenoreceptor agonist.^[2]^[3]^[4]^[5]^[6] Adrenergic receptors, or adrenoceptors, belong to the class of G protein–coupled receptors, and are the most prominent receptors in the adipose membrane, besides also being expressed in skeletal muscle tissue. These adipose membrane receptors are classified as either α or β adrenoceptors. Although these adrenoceptors share the same messenger, cyclic adenosine monophosphate (cAMP), the specific transduction pathway depends on the receptor type (α or β). Higenamine partly exerts its actions by the activation of an enzyme,adenylate cyclase, responsible for boosting the cellular concentrations of the adrenergic second messenger, cAMP.^[7]

In a rodent model, it was found that higenamine produced cardiotonic, vascular relaxation, and bronchodilator effects.^[8]^[9] In particular, higenamine, via a beta-adrenoceptor mechanism, induced relaxation in rat corpus cavernosum, leading to improved vasodilation and erectile function.

Related to improved vasodilatory signals, higenamine has been shown in animal models to possess antiplatelet and antithrombotic activity via a cAMP-dependent pathway, suggesting higenamine may contribute to enhanced vasodilation and arterial integrity.^[2]^[7]^[9]^[10]

Toxicity

Regarding toxicity, researchers have suggested that the levels of higenamine reported in food consumption (estimated 47.5 mg in a 9-ounce serving of Lotus) would be comparable to the amount used in food supplements.^{[citation needed]} Higenamine is a beta-adrenergic agonist which has effects on the function of trachea and heart muscles.^[11]^[12]During a study of acute toxicity, mice were orally administered the compound at a dose of 2 g per kg of bodyweight. No mice died during the study.^[13] higenamine is one of the main chemicals in a plant called aconite. Aconite has been shown to cause serious heart-related side effects including arrhythmias and even death. in some sources of HIGENAMINE from certain plants that have Aconite

PAPER

Chemical & Pharmaceutical Bulletin (1978), 26(7), 2284-5

https://www.jstage.jst.go.jp/article/cpb1958/26/7/26_7_2284/_pdf

PATENT

CN 103554022

http://google.com/patents/CN103554022B?cl=en

Example 1:

[0024] to the S-necked flask 200mL of anhydrous ammonia clever four furans, lOg instrument crumbs, olive mix was added 0. 5g ship, continue to embrace the mix was added 10 minutes after which 2 drops of 1,2-B burning desert, Continue mixing until the reaction mixture embrace color disappeared, the reaction was cooled to square ° C, and slowly mixed solution thereto 31. 6g4- methoxy Desert Festival and 50mL tetraammine clever furans dropped, about 60min addition was complete, the reaction fluid continues to cool to -65 ° C, to which was slowly dropping 20 percent, 7-dimethoxy-3,4-diamine different wow beep and a mixed solution of ammonia lOOmL four clever furans, the addition was complete continue to maintain – 65 ° C for 2 hours after the embrace slowly warmed 0 ° C, maintaining the internal temperature of 100 ° C 〇 blood slowly added to the reaction mixture, the addition was completed adding 200 blood continues to embrace mixed with ethyl acetate after 0.5 hours, allowed to stand liquid separation, organic phase was separated, dried over anhydrous sulfate steel, concentrated to afford 6, 7-dimethoxy -l- (4- methoxy section yl) -1,2, 3, 4-isopropyl tetraammine wow toot 24. 9g, a yield of 76.1%.

[00 Qiao] to the reaction flask prepared above 6, 7-dimethoxy -l- (4- methoxybenzyl) -1,2, 3, 4 tetraammine different wow beep 24. 9g , 47% aqueous ammonia desert 200 blood acid heated to 130 ° C reflux of cooled to room temperature, precipitation of large amount of solid, filtered higenamine ammonia salt desert, the solid was added 1. of water and continue to add 50 Blood mixed with ammonia football ground, filtered higenamine to higenamine was added lL4mol / L aqueous hydrochloric acid, 80 ° C heat to embrace mixed, cooled to 25 ° C filtration and drying to obtain a final product hydrochloric acid higenamine 11. 7g, a yield of 73.3%.

References

http://ec.europa.eu/food/food/biotechnology/novelfood/novel_food_catalogue_en.htm
Tsukiyama, M; Ueki, T; Yasuda, Y; Kikuchi, H; Akaishi, T; Okumura, H; Abe, K (2009). “Beta2-adrenoceptor-mediated tracheal relaxation induced by higenamine from Nandina domestica Thunberg”. Planta Medica 75 (13): 1393–9. doi:10.1055/s-0029-1185743. PMID 19468973.
Kashiwada, Y; Aoshima, A; Ikeshiro, Y; Chen, YP; Furukawa, H; Itoigawa, M; Fujioka, T; Mihashi, K; et al. (2005). “Anti-HIV benzylisoquinoline alkaloids and flavonoids from the leaves of Nelumbo nucifera, and structure-activity correlations with related alkaloids”.Bioorganic & Medicinal Chemistry 13 (2): 443–8. doi:10.1016/j.bmc.2004.10.020.PMID 15598565.
Kimura, I; Chui, LH; Fujitani, K; Kikuchi, T; Kimura, M (1989). “Inotropic effects of (+/-)-higenamine and its chemically related components, (+)-R-coclaurine and (+)-S-reticuline, contained in the traditional sino-Japanese medicines “bushi” and “shin-i” in isolated guinea pig papillary muscle”. Japanese journal of pharmacology 50 (1): 75–8.doi:10.1254/jjp.50.75. PMID 2724702.
Kang, YJ; Lee, YS; Lee, GW; Lee, DH; Ryu, JC; Yun-Choi, HS; Chang, KC (1999). “Inhibition of activation of nuclear factor kappaB is responsible for inhibition of inducible nitric oxide synthase expression by higenamine, an active component of aconite root”. The Journal of Pharmacology and Experimental Therapeutics 291 (1): 314–20.PMID 10490919.
Yun-Choi, HS; Pyo, MK; Park, KM; Chang, KC; Lee, DH (2001). “Anti-thrombotic effects of higenamine”. Planta Medica 67 (7): 619–22. doi:10.1055/s-2001-17361.PMID 11582538.
Kam, SC; Do, JM; Choi, JH; Jeon, BT; Roh, GS; Chang, KC; Hyun, JS (2012). “The relaxation effect and mechanism of action of higenamine in the rat corpus cavernosum”.International Journal of Impotence Research 24 (2): 77–83. doi:10.1038/ijir.2011.48.PMID 21956762.
Bai, G; Yang, Y; Shi, Q; Liu, Z; Zhang, Q; Zhu, YY (2008). “Identification of higenamine in Radix Aconiti Lateralis Preparata as a beta2-adrenergic receptor agonist1”. Acta pharmacologica Sinica 29 (10): 1187–94. doi:10.1111/j.1745-7254.2008.00859.x.PMID 18817623.
Pyo, MK; Lee, DH; Kim, DH; Lee, JH; Moon, JC; Chang, KC; Yun-Choi, HS (2008). “Enantioselective synthesis of (R)-(+)- and (S)-(-)-higenamine and their analogues with effects on platelet aggregation and experimental animal model of disseminated intravascular coagulation”. Bioorganic & Medicinal Chemistry Letters 18 (14): 4110–4.doi:10.1016/j.bmcl.2008.05.094. PMID 18556200.
Liu, W; Sato, Y; Hosoda, Y; Hirasawa, K; Hanai, H (2000). “Effects of higenamine on regulation of ion transport in guinea pig distal colon”. Japanese journal of pharmacology 84(3): 244–51. doi:10.1254/jjp.84.244. PMID 11138724.
Wong, KK; Lo, CF; Chen, CM (1997). “Endothelium-dependent higenamine-induced aortic relaxation in isolated rat aorta”. Planta Medica 63 (2): 130–2. doi:10.1055/s-2006-957628. PMID 9140225.
Ueki, T; Akaishi, T; Okumura, H; Morioka, T; Abe, K (2011). “Biphasic tracheal relaxation induced by higenamine and nantenine from Nandina domestica Thunberg”. Journal of pharmacological sciences 115 (2): 254–7. doi:10.1254/jphs.10251sc. PMID 21282929.
Lo, CF; Chen, CM (1997). “Acute toxicity of higenamine in mice”. Planta Medica 63 (1): 95–6. doi:10.1055/s-2006-957619. PMID 9063102.

banned in ncaa https://www.ncaa.org/sites/default/files/2015-16%20NCAA%20Banned%20Drugs.pdf

CN1539823A *	Oct 27, 2003	Oct 27, 2004	中国医学科学院药物研究所	Method for preparing new demethyl conclaurine and medinal salt
CN1764647A *	Mar 23, 2004	Apr 26, 2006	埃科特莱茵药品有限公司	Tetrahydroisoquinolyl acetamide derivatives for use as orexin receptor antagonists
CN103351338A *	Jun 17, 2013	Oct 16, 2013	张家港威胜生物医药有限公司	Simple preparation process of higenamine hydrochloride
US20060030586 *	Sep 27, 2004	Feb 9, 2006	Education Center Of Traditional Chinese Medicine Co.	Method and health food for preventing and/or alleviating psychiatric disorder, and/or for effectuating sedation
WO2011038169A2 *	Sep 24, 2010	Mar 31, 2011	Mallinckrodt Inc.	One-pot preparation of hexahydroisoquinolines from amides

Higenamine
Names

IUPAC name 1-[(4-Hydroxyphenyl)methyl]-1,2,3,4-tetrahydroisoquinoline-6,7-diol
Other names norcoclaurine, demethylcoclaurine
Identifiers
CAS Number	5843-65-2 106032-53-5 (R) 22672-77-1 (S)
ChEBI	CHEBI:18418
ChEMBL	ChEMBL19344
ChemSpider	102800
Jmol 3D model	Interactive image
KEGG	C06346
MeSH	higenamine
PubChem	114840
InChI [show]
SMILES [show]
Properties
Chemical formula	C₁₆H₁₇NO₃
Molar mass	271.32 g·mol⁻¹

/////

Chelation-Controlled Bergman Cyclization: Synthesis and Reactivity of Enediynyl Ligands

SYNTHESIS, Uncategorized Comments Off

May 232016

Chelation-Controlled Bergman Cyclization: Synthesis and Reactivity of Enediynyl Ligands

● Basak, Amit; Mandal, Subrata; Bag, Subhendu Sekhar●

Chemical Reviews2003, 103(10), 4077-4094.

Abstract: A review with 150 references.

see

http://pubs.acs.org/doi/abs/10.1021/cr020069k

Dr. SUBHENDU SEKHAR BAG

Associate Professor

Bioorganic Chemistry Laboratory

Room No. CHF-208 (O); CH-103 (Lab.); Core-2

Department of Chemistry

Indian Institute of Technology Guwhati,

Guwahati-781 039, Assam, INDIA.

Ph : +91-361-258-2324 (O);

+91-361-258-4324 (R)

Mobile: 0361-258-4324

Fax: +91-361-258-2349

Email: ssbag75@iitg.ernet.in//ssbag75@yahoo.co.in

////////////Bergman Cyclization, Enediynyl Ligands

DSM 265 a promising Antimalarial

phase 2, Uncategorized Comments Off

May 232016

DSM265

DSM-265; PfSPZ

2-(1,1-difluoroethyl)-5-methyl-N-(4-(pentafluoro-l6-sulfanyl)phenyl)-[1,2,4]triazolo[1,5-a]pyrimidin-7-amine

2-(l,l-difluoroethyl)-5-methyl-N-[4-(pentafluoro- ⁶– sulfanyl)phenyl] [ 1 ,2,4]triazolo[ 1 ,5-a]pyrimidin-7-amine.

(OC-6-21)-[4-[[2-(1,1-Difluoroethyl)-5-methyl[1,2,4]triazolo[1,5-a]pyrimidin-7-yl]amino]phenyl]pentafluorosulfur

1282041-94-4
Chemical Formula: C14H12F7N5S
Exact Mass: 415.0702

Board Of Regents, University Of Texas System, Monash University, Medicines For Malaria Venture

DSM265 is a long-duration, potent and selective dihydroorotate dehydrogenase (DHODH)) inhibitor. DSM265 is potential useful for the prevention and treatment of malaria. DSM265 is the first DHODH inhibitor to reach clinical development for treatment of malaria. DSM265 is highly selective toward DHODH of the malaria parasite Plasmodium, efficacious against both blood and liver stages of P. falciparum, and active against drug-resistant parasite isolates. DSM265 has advantages over current treatment options that are dosed daily or are inactive against the parasite liver stage.

OriginatorMonash University; University of Texas Southwestern Medical Center; University of Washington
Developer Center for Infectious Disease Research; Fred Hutchinson Cancer Research Center; Medicines for Malaria Venture; Takeda; United States Department of Defense
Class Antimalarials; Pyrimidines; Small molecules; Triazoles
Mechanism of Action Dihydroorotate dehydrogenase inhibitors

Phase II Malaria
Phase I Malaria

Most Recent Events

25 Apr 2016 Medicines for Malaria Venture and AbbVie plan a phase I bioavailability trial in Healthy volunteers in USA (PO, Granule) (NCT02750384)
01 Mar 2016 Phase-I clinical trials in Malaria prevention (In volunteers) in USA (PO) (NCT02562872)
01 Jan 2016 Phase-II clinical trials in Malaria in Peru (PO) (NCT02123290)

Malaria is one of the most significant causes of childhood mortality, but disease control efforts are threatened by resistance of the Plasmodium parasite to current therapies. Continued progress in combating malaria requires development of new, easy to administer drug combinations with broad-ranging activity against all manifestations of the disease. DSM265, a triazolopyrimidine-based inhibitor of the pyrimidine biosynthetic enzyme dihydroorotate dehydrogenase (DHODH), is the first DHODH inhibitor to reach clinical development for treatment of malaria. We describe studies profiling the biological activity, pharmacological and pharmacokinetic properties, and safety of DSM265, which supported its advancement to human trials. DSM265 is highly selective toward DHODH of the malaria parasite Plasmodium, efficacious against both blood and liver stages of P. falciparum, and active against drug-resistant parasite isolates. Favorable pharmacokinetic properties of DSM265 are predicted to provide therapeutic concentrations for more than 8 days after a single oral dose in the range of 200 to 400 mg. DSM265 was well tolerated in repeat-dose and cardiovascular safety studies in mice and dogs, was not mutagenic, and was inactive against panels of human enzymes/receptors. The excellent safety profile, blood- and liver-stage activity, and predicted long half-life in humans position DSM265 as a new potential drug combination partner for either single-dose treatment or once-weekly chemoprevention. DSM265 has advantages over current treatment options that are dosed daily or are inactive against the parasite liver stage.

A new single-dose malaria drug is offering promise as both a cure to malaria and also a way to prevent the disease according to researchers at UT Southwestern Medical Center. The new drug, which is known as DSM265, kills the drug-resistant malaria parasites in the blood and liver by targeting the ability of the parasites to replicate.

malaria

Malaria is a very infectious disease that is transmitted by mosquitoes, and it kills about 600,000 people worldwide every year. Most of the people who are killed by malaria are under 5-years-old, and it’s more common in sub-Saharan Africa. Almost 200 million cases of malaria are reported every year, with about 3 billion people in 97 countries at risk for the disease. Lead author Dr. Margaret Phillips, who is a professor of Pharmacology at UT Southwestern said that this could be the first single-dose cure for malaria, and would be used in partnership with another drug. This drug could also be developed into a once-a-week preventive vaccination as well, and the results of the study were just published in Science Translational Medicine. Not only was UT Southwestern involved in the research study, but Monash Institute of Pharmaceutical Sciences in Australia, the University of Washington, and the not-for-profit Medicines for Malaria Venture was also involved.

Malaria is one of the most deadly infectious diseases in human history with 3.2 billion people in 97 countries at risk. An estimated 444,000 deaths from malaria were reported by the WHO in 2015 and ∼90% of these occurred in sub-Saharan Africa, mostly among children under the age of five. Human malaria, which is transmitted by the female Anopheles mosquito, can be caused by five species of Plasmodia; however, Plasmodium falciparum and Plasmodium vivax are the most signficant.P. falciparum is dominant in Africa and accounts for most of the deaths, while P. vivax has a larger global distribution.

To simplify treatment options it is desirable that new drugs be efficacious against all human infective species. Malaria is a treatable disease and malarial control programs depend on drug therapy for treatment and chemoprevention, and on insecticides (including insecticide impregnated bed nets) to prevent transmission.

A large collection of drugs has been used for the treatment of malaria, but many of the most important compounds have been lost to drug resistance (e.g., chloroquine and pyrimethamine).Artemisinin combination therapies (ACT) replaced older treatments, becoming highly effective, crucial tools in global efforts that have led to the decline in malaria deaths over the past decade. However, resistance to the artemisinin components (associated with Kelch13 propeller protein mutations has been found in Southeast Asia putting at risk malaria treatment programs. To combat drug resistance a significant effort is underway to identify new compounds that can be used for the treatment of malaria, with several new entities currently in clinical development.

The triazolopyrimidine DSM265 developed by the group is the first antimalarial agent that targets dihydroorotate dehydrogenase (DHODH) to reach clinical development, validating this target for the treatment of malaria. DHODH is a mitochondrial enzyme that is required for the fourth step of de novo pyrimidine biosynthesis, catalyzing the flavin-dependent oxidation of dihydroorotate to orotic acid with mitochondrially derived coenzyme Q (CoQ) serving as a second substrate. Pyrimidines are essential for both RNA and DNA biosynthesis, and because Plasmodia do not encode pyrimidine salvage enzymes, which are found in humans and other organisms, the de novo pyrimidine pathway and DHODH are essential to the parasite.

They identified the triazolopyrimidine DHODH inhibitor series by a target-based high throughput screen, and the initial lead DSM1 (2) was shown to selectively inhibit P. falciparumDHODH and to kill parasites in vitro, but it was ineffective in vivo due to poor metabolic properties. The series was subsequently optimized to improve its in vivo properties resulting in the identification of DSM74 (3), which while metabolically stable lacked potencyX-ray structures of 2 and 3 bound to PfDHODH were then used to guide the medicinal chemistry program in the search for more potent analogues, resulting in the identification of 1.

SYNTHESIS

PATENT

WO 2015165660

PAPER

Journal of Medicinal Chemistry (2012), 55(17)

http://pubs.acs.org/doi/abs/10.1021/jm300351w

Plasmodium falciparum causes approximately 1 million deaths annually. However, increasing resistance imposes a continuous threat to existing drug therapies. We previously reported a number of potent and selective triazolopyrimidine-based inhibitors of P. falciparum dihydroorotate dehydrogenase that inhibit parasite in vitro growth with similar activity. Lead optimization of this series led to the recent identification of a preclinical candidate, showing good activity against P. falciparum in mice. As part of a backup program around this scaffold, we explored heteroatom rearrangement and substitution in the triazolopyrimidine ring and have identified several other ring configurations that are active as PfDHODH inhibitors. The imidazo[1,2-a]pyrimidines were shown to bind somewhat more potently than the triazolopyrimidines depending on the nature of the amino aniline substitution. DSM151, the best candidate in this series, binds with 4-fold better affinity (PfDHODH IC₅₀ = 0.077 μM) than the equivalent triazolopyrimidine and suppresses parasites in vivo in the Plasmodium berghei model.

PATENT

WO 2011041304

Scheme 3

Example 44: Synthesis of 2-(l,l-difluoroethyl)-5-methyl-N-[4-(pentafluoro- ⁶– sulfanyl)phenyl] [ 1 ,2,4]triazolo[ 1 ,5-a]pyrimidin-7-amine.

A suspension of Intermediate 3 (5.84 g, 25.09 mmol) and 4-aminophenylsulfur pentafluoride (MANCHESTER, 5.5 g, 25.09 mmol) in ethanol (150 mL) was heated at 50 °C for 1 h. Heating resulted in the precipitation of a solid. The reaction mixture was concentrated under vacuum, redissolved in DCM (300 mL) and washed with aq. Na₂C0₃ (2 x 350 mL). The organic layer was dried over Na₂S0₄ and filtered. Then 8 g of silica gel were added and the mixture was concentrated under vacuum to dryness. The residue was purified (silica gel column, eluting with Hexane/EtOAc mixtures from 100:0 to 50:50%) to afford the title compound as a white solid.

1H NMR (400 MHz, DMSO-d₆) δ ppm: 10.60 (bs, 1H), 7.97 (d, 2H), 7.67 (d, 2H), 6.79 (s, 1H), 2.47 (s, 3H), 2.13 (t, 3H); [ES+ MS] m/z 416 (MH)⁺.

PAPER

Journal of Medicinal Chemistry (2011), 54(15), 5540-5561

http://pubs.acs.org/doi/abs/10.1021/jm200592f

Drug therapy is the mainstay of antimalarial therapy, yet current drugs are threatened by the development of resistance. In an effort to identify new potential antimalarials, we have undertaken a lead optimization program around our previously identified triazolopyrimidine-based series of Plasmodium falciparum dihydroorotate dehydrogenase (PfDHODH) inhibitors. The X-ray structure of PfDHODH was used to inform the medicinal chemistry program allowing the identification of a potent and selective inhibitor (DSM265) that acts through DHODH inhibition to kill both sensitive and drug resistant strains of the parasite. This compound has similar potency to chloroquine in the humanized SCID mouse P. falciparum model, can be synthesized by a simple route, and rodent pharmacokinetic studies demonstrated it has excellent oral bioavailability, a long half-life and low clearance. These studies have identified the first candidate in the triazolopyrimidine series to meet previously established progression criteria for efficacy and ADME properties, justifying further development of this compound toward clinical candidate statu

PAPER

Malaria persists as one of the most devastating global infectious diseases. The pyrimidine biosynthetic enzyme dihydroorotate dehydrogenase (DHODH) has been identified as a new malaria drug target, and a triazolopyrimidine-based DHODH inhibitor 1 (DSM265) is in clinical development. We sought to identify compounds with higher potency against PlasmodiumDHODH while showing greater selectivity toward animal DHODHs. Herein we describe a series of novel triazolopyrimidines wherein the p-SF₅-aniline was replaced with substituted 1,2,3,4-tetrahydro-2-naphthyl or 2-indanyl amines. These compounds showed strong species selectivity, and several highly potent tetrahydro-2-naphthyl derivatives were identified. Compounds with halogen substitutions displayed sustained plasma levels after oral dosing in rodents leading to efficacy in the P. falciparum SCID mouse malaria model. These data suggest that tetrahydro-2-naphthyl derivatives have the potential to be efficacious for the treatment of malaria, but due to higher metabolic clearance than 1, they most likely would need to be part of a multidose regimen

Tetrahydro-2-naphthyl and 2-Indanyl Triazolopyrimidines TargetingPlasmodium falciparum Dihydroorotate Dehydrogenase Display Potent and Selective Antimalarial Activity

Sreekanth Kokkonda^†, Xiaoyi Deng‡, Karen L. White^§, Jose M. Coteron^∥, Maria Marco^∥, Laura de las Heras^∥,John White^†, Farah El Mazouni‡, Diana R. Tomchick⊥, Krishne Manjalanagara^#, Kakali Rani Rudra^#, Gong Chen^§,Julia Morizzi^§, Eileen Ryan^§, Werner Kaminsky^†, Didier Leroy^∇, María Santos Martínez-Martínez^∥, Maria Belen Jimenez-Diaz^∥, Santiago Ferrer Bazaga^∥, Iñigo Angulo-Barturen^∥, David Waterson^∇, Jeremy N. Burrows^∇, Dave Matthews^∇, Susan A. Charman^§, Margaret A. Phillips^*‡, and Pradipsinh K. Rathod^*^†

^† Departments of Chemistry and Global Health, University of Washington, Seattle, Washington 98195, United States

^‡Departments of Pharmacology and ^⊥Biophysics, University of Texas Southwestern Medical Center at Dallas, 6001 Forest Park Blvd, Dallas, Texas 75390-9041, United States

^§ Centre for Drug Candidate Optimisation, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia

^∥ GSK, Tres Cantos Medicines Development Campus, Severo Ochoa, Madrid 28760 Spain

^# Syngene International Ltd., Bangalore 560 099, India

^∇ Medicines for Malaria Venture, 1215 Geneva, Switzerland

J. Med. Chem., Article ASAP

DOI: 10.1021/acs.jmedchem.6b00275

http://pubs.acs.org/doi/full/10.1021/acs.jmedchem.6b00275

*Phone: 214-645-6164. E-mail: margaret.phillips@UTSouthwestern.edu., *Phone: 206-221-6069. E-mail:rathod@chem.washington.edu.

REFERENCES

1: Phillips MA, Lotharius J, Marsh K, White J, Dayan A, White KL, Njoroge JW, El
Mazouni F, Lao Y, Kokkonda S, Tomchick DR, Deng X, Laird T, Bhatia SN, March S,
Ng CL, Fidock DA, Wittlin S, Lafuente-Monasterio M, Benito FJ, Alonso LM,
Martinez MS, Jimenez-Diaz MB, Bazaga SF, Angulo-Barturen I, Haselden JN, Louttit
J, Cui Y, Sridhar A, Zeeman AM, Kocken C, Sauerwein R, Dechering K, Avery VM,
Duffy S, Delves M, Sinden R, Ruecker A, Wickham KS, Rochford R, Gahagen J, Iyer
L, Riccio E, Mirsalis J, Bathhurst I, Rueckle T, Ding X, Campo B, Leroy D, Rogers
MJ, Rathod PK, Burrows JN, Charman SA. A long-duration dihydroorotate
dehydrogenase inhibitor (DSM265) for prevention and treatment of malaria. Sci
Transl Med. 2015 Jul 15;7(296):296ra111. doi: 10.1126/scitranslmed.aaa6645.
PubMed PMID: 26180101; PubMed Central PMCID: PMC4539048.

2: Held J, Jeyaraj S, Kreidenweiss A. Antimalarial compounds in Phase II clinical
development. Expert Opin Investig Drugs. 2015 Mar;24(3):363-82. doi:
10.1517/13543784.2015.1000483. Epub 2015 Jan 7. Review. PubMed PMID: 25563531.

3: Gamo FJ. Antimalarial drug resistance: new treatments options for Plasmodium.
Drug Discov Today Technol. 2014 Mar;11:81-88. doi: 10.1016/j.ddtec.2014.03.002.
Review. PubMed PMID: 24847657.

4: Coteron JM, Marco M, Esquivias J, Deng X, White KL, White J, Koltun M, El
Mazouni F, Kokkonda S, Katneni K, Bhamidipati R, Shackleford DM, Angulo-Barturen
I, Ferrer SB, Jiménez-Díaz MB, Gamo FJ, Goldsmith EJ, Charman WN, Bathurst I,
Floyd D, Matthews D, Burrows JN, Rathod PK, Charman SA, Phillips MA.
Structure-guided lead optimization of triazolopyrimidine-ring substituents
identifies potent Plasmodium falciparum dihydroorotate dehydrogenase inhibitors
with clinical candidate potential. J Med Chem. 2011 Aug 11;54(15):5540-61. doi:
10.1021/jm200592f. Epub 2011 Jul 14. PubMed PMID: 21696174; PubMed Central PMCID:
PMC3156099.