WO2019051465A1 - Rad51 inhibitors - Google Patents

Rad51 inhibitors Download PDF

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Publication number
WO2019051465A1
WO2019051465A1 PCT/US2018/050391 US2018050391W WO2019051465A1 WO 2019051465 A1 WO2019051465 A1 WO 2019051465A1 US 2018050391 W US2018050391 W US 2018050391W WO 2019051465 A1 WO2019051465 A1 WO 2019051465A1
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Prior art keywords
compound
cancer
phenyl
pharmaceutically acceptable
monocyclic
Prior art date
Application number
PCT/US2018/050391
Other languages
French (fr)
Inventor
Alfredo C. Castro
Casey Cameron Mccomas
Joseph Vacca
Tyler MACLAY
Original Assignee
Cyteir Therapeutics, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to MX2020002745A priority Critical patent/MX2020002745A/en
Priority to IL293332A priority patent/IL293332B2/en
Priority to CN201880072664.5A priority patent/CN111542521B/en
Priority to EP18782257.2A priority patent/EP3681884B1/en
Priority to BR112020004828-3A priority patent/BR112020004828A2/en
Priority to SI201830740T priority patent/SI3681884T1/en
Priority to CA3075062A priority patent/CA3075062A1/en
Application filed by Cyteir Therapeutics, Inc. filed Critical Cyteir Therapeutics, Inc.
Priority to PL18782257.2T priority patent/PL3681884T3/en
Priority to ES18782257T priority patent/ES2925218T3/en
Priority to JP2020514206A priority patent/JP7265537B2/en
Priority to KR1020207010585A priority patent/KR102718671B1/en
Priority to IL273156A priority patent/IL273156B/en
Priority to RU2020113064A priority patent/RU2795882C2/en
Priority to SG11202002069WA priority patent/SG11202002069WA/en
Priority to AU2018328818A priority patent/AU2018328818C1/en
Priority to EP22175891.5A priority patent/EP4112616A1/en
Priority to DK18782257.2T priority patent/DK3681884T3/en
Publication of WO2019051465A1 publication Critical patent/WO2019051465A1/en
Priority to PH12020550079A priority patent/PH12020550079A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/426—1,3-Thiazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
    • C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
    • C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D277/22—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
    • C07D277/28—Radicals substituted by nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
    • C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
    • C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D277/22—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
    • C07D277/30—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
    • C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
    • C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D277/32—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D277/38—Nitrogen atoms
    • C07D277/42—Amino or imino radicals substituted by hydrocarbon or substituted hydrocarbon radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • This application is directed to inhibitors of RAD51, and methods for their use, such as to treat conditions including cancers, autoimmune diseases, immune deficiencies, and neurodegenerative diseases.
  • RAD51 is a eukaryote gene.
  • the protein encoded by this gene is a member of the RAD51 protein family which assists in repair of DNA double strand breaks.
  • RAD51 family members are homologous to the bacterial RecA, Archaeal RadA and yeast RAD51. The protein is highly conserved in most eukaryotes, from yeast to humans.
  • RAD51 is a 339-amino acid protein that plays a major role in homologous recombination of DNA during double strand break (DSB) repair.
  • RAD51 catalyzes strand transfer between a broken sequence and its undamaged homologue to allow re-synthesis of the damaged region (see homologous recombination models).
  • the RAD51 inhibitors of the present invention inhibit homologous recombination by altering the nucleocytoplasmic distribution of RAD51 following DNA damage induction.
  • the RAD51 inhibitors of the present invention reduce the repair of AID-induced DNA double strand breaks, leading to AID-dependent cytotoxicity in both normal and malignant B -lymphocytes.
  • Certain of these RAD51 inhibitors have superior cell permeability as measured in Caco-2 cells (see Example 76).
  • the RAD51 inhibitors with good cell permeability several have superior metabolic stability (as measured by a liver microsome assay, see Example 77) and exposure, including oral exposure (see Example 79).
  • the present invention provides a compound represented by Structural Formula I:
  • the present invention also provides a pharmaceutical composition comprising a compound as described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or diluent.
  • the present invention further provides a method of treating a cancer, an autoimmune disease, an immune deficiency, or a neurodegenerative disease.
  • the method comprises administering to a subject in need thereof an effective amount of a compound of disclosed herein or a pharmaceutically acceptable salt thereof or a pharmaceutical composition disclosed herein.
  • a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical compositions disclosed herein for the preparation of a medicament for the treatment of a cancer, an autoimmune disease, an immune deficiency, or a neurodegenerative disease.
  • the invention provides a compound represented by Structural Formula I: I;
  • the thiazole ring is optionally substituted with -F or -CI;
  • Cy is -(C3-C 7 )cycloalkyl, bridged (C6-C 12 ) cycloalkyl, or a 4-12 membered heterocyclic ring, each of which is optionally substituted with one or more groups selected from the group consisting of halogen, -OH, (Q-G alkyl, and
  • X 5 when X 5 is connected with a carbon ring atom of Cy, X 5 is NR or O; X 6 is NR a or O;
  • R 1 is (Ci-C 5 )alkyl
  • R 3 is (Ci-C 5 )alkyl, -CH 2 -phenyl, -(C 3 -C 7 )cycloalkyl, -CH 2 -(C 3 -C 7 )cycloalkyl, -CH 2 -monocyclic 3-7 membered heterocyclic ring, or monocyclic 3-7 membered heterocyclic ring, wherein the (Ci-C5)alkyl, -(C 3 -C 7 )cycloalkyl, phenyl or monocyclic 3-7 membered heterocyclic ring represented by R 3 or in the group represented by R 3 is optionally substituted with one or more groups selected from the group consisting of halogen, -OH, (Ci-C 4 )alkyl, halomethyl, halomethoxy, -CN, and (Ci-C 4 )alkoxy;
  • R 2 is -NR a C(0)0(Ci-C 4 )alkyl; -NR a C(0)NR a (Ci-C 4 )alkyl; -NR a C(0)0(C 2 - C 4 )alkenyl;
  • the (Ci-C 4 )alkyl and the (C 2 -C 4 )alkenyl in the group represented by R are each optionally and independently substituted with one or more groups selected from the group consisting of halogen, N 3 , -OR , -NR R , -(C 3 -C6)cycloalkyl, phenyl, a monocyclic 3- 7 membered heterocyclic ring, and a monocyclic 5-6 membered heteroaromatic ring;
  • phenyl in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen, -CH 3 , halomethyl,
  • heteroaromatic ring in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen, -CN, - CH 3 , halomethyl, halomethoxy, -OR a and -NR a R a ; and
  • each R is independently -H or -CH 3 .
  • the invention provides a compound represented by Structural Formula II:
  • the thiazole ring is optionally substituted with -F or -CI;
  • Cy is cyclohexyl or a 6-membered monocyclic heterocyclic ring
  • X 5 and X 6 are each independently NR or O;
  • R 1 is (Ci-C 5 )alkyl
  • R is (Ci-C5)alkyl or monocyclic 3-7-membered heterocyclic ring
  • R 2 is -NR a C(0)0(Ci-C 4 )alkyl; -NR a C(0)NR a (Ci-C 4 )alkyl; -NR a C(0)0(C 2 - C 4 )alkenyl;
  • (Ci-C 4 )alkyl and the (C 2 -C 4 )alkenyl in the group represented by R are each optionally and independently substituted with one or more halogen, N 3 , -OR , -NR R , -(C 3 -C 6 )cycloalkyl, phenyl, monocyclic 3-7-membered heterocyclic ring, or monocyclic 5-6-membered heteroaromatic ring;
  • -(C 3 -C 6 )cycloalkyl in the group represented by R is optionally substituted with one or more halogen, -CH 3 , -OR or -NR R ;
  • phenyl in the group represented by R is optionally substituted with one or more halogen, -CH 3 , halomethyl, halomethoxy, -OR , or -N 3 ;
  • heteroaromatic ring in the group represented by R is optionally substituted with one or more halogen, -CH 3 , halomethyl, halomethoxy, -OR or -NR R ; and each R is independently -H or -CH 3 .
  • the invention provides a compound according to Structural Formula I, or a pharmaceutically acceptable salt thereof, wherein Cy is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl; azetidinyl, azepanyl, diazaspiro[4.4]nonyl, diazaspiro[3.5]nonyl, diazepanyl, dihydroimidazole, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, dihydropyrimidinyl, dihydrothienyl, dihydrothiophenyl,
  • dihydrothiopyranyl hexahydropyridazinyl, hexahydropyrimidinyl, hydantoinyl, indolinyl, isoindolinyl, morpholinyl, oxiranyl, oxetanyl, piperidinyl, piperazinyl, pyrrolidinonyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydroimidazole, tetrahydroindolyl, tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridinyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, thio morpholinyl, tropanyl, valerolactamyl; bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bi
  • the invention provides a compound according to Structural Formula I or II, or a pharmaceutically acceptable salt thereof, wherein Cy is cyclohexyl, morpholinyl, thiomorpholinyl, piperidinyl, piperazinyl, hexahydropyridazinyl,
  • the invention provides a compound represented by Structural Formula III,
  • X 7 is NH or O;
  • R 4 is (Ci-C 4 )alkyl, (C 3 -C 6 )cycloalkyl, or a monocyclic 3-7 membered heterocyclic ring;
  • phenyl in the group represented by R 4 is optionally substituted with one or more groups selected from the group consisting of halogen, -CH 3 , halomethyl,
  • heteroaromatic ring in the group represented by R 4 is optionally substituted with one or more groups selected from the group consisting of halogen and -CH 3 ; and the remaining variables are as defined in the first, second, third, and/or fourth
  • the invention provides a compound according to Structural
  • the invention provides a compound represented by
  • the invention provides a compound represented by Structural Formula V,
  • the invention provides a compound represented by Structural Formula VI:
  • the invention provides a compound represented by Structural Formula VII:
  • the invention provides a compound represented by
  • the invention provides a compound represented by
  • the invention provides a compound according to
  • the invention provides a compound according to
  • Structural Formula I, II, or III, or a pharmaceutically acceptable salt thereof wherein Cy is l,7-diazaspiro[4.4]nonyl, 2,7-diazaspiro[4.4]nonyl, 2,7-diazaspiro[3.5]nonyl, 1,4-diazepanyl, 2,5-diazabicyclo[2.2. l]heptanyl, 3,8-diazabicyclo[3.2.
  • the invention provides a compound according to Structural Formula III, IV, V, VI, VII, VIII, or IX, or a pharmaceutically acceptable salt thereof, wherein R 4 is -(Ci-C 3 )alkyl, (C 3 -C6)cycloalkyl, or a monocyclic 3-7 membered heterocyclic ring, wherein the -(Ci-C 3 )alkyl is optionally substituted with (i) phenyl optionally substituted by one or more halogen or -CH 3 ; (ii) a monocyclic 5-6 membered heteroaromatic ring optionally substituted by one or more halogen or -CH 3 ; or (iii) a monocyclic 3-7 membered heterocyclic ring optionally substituted by one or more groups selected from the group consisting of halogen and -CH 3 ; and the remaining variables are as defined in the first, second, third, fourth, fifth, seventh, eighth, ninth, tenth,
  • the invention provides a compound according to
  • R 4 is -(Ci-C 3 )alkyl, -CHR -phenyl, -CHR -5-6 membered heteraromatic ring, or -CHR -3-7 membered monocyclic heterocyclic ring, wherein the phenyl, 5-6 membered heteraromatic ring or 3-7 membered monocyclic heterocyclic ring in the group represented by R 4 is optionally substituted one or more groups selected from the group consisting of halogen and -CH 3 ; and the remaining variables are as defined in the first, second, third, fourth, fifth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, and/or fourteenth embodiments.
  • the invention provides a compound according to
  • R is (Q-G alkyl, -(C4-C6)cycloalkyl, -CH 2 -phenyl, -CH 2 -monocyclic 4-6 membered heterocyclic ring, or monocyclic 4-6 membered heterocyclic ring, wherein the phenyl or monocyclic 4-6 membered heterocyclic ring represented by R or in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen,
  • the invention provides a compound represented by Structural Formula X:
  • variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenth embodiments.
  • the invention provides a compound represented by Structural Formula XI:
  • variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenth embodiments.
  • the invention provides a compound represented by Structural Formula XII:
  • the invention provides a compound represented by Structural Formula XIII(a) or XIII(b):
  • variables are as defined in the first, second, third, fourth, fifth, sixth, eighth, fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenth embodiments.
  • the invention provides a compound represented by Structural Formula XI
  • variables are as defined in the first, second, third, fourth, fifth, sixth, tenth, fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenth embodiments.
  • the invention provides a compound according to
  • R 3 is isopropyl or oxetanyl. In another alternative embodiment, R 3 is isopropyl.
  • the invention provides a compound according to Structural Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII(a), XIII(b), XIV, or a pharmaceutically acceptable salt thereof, wherein R 1 is tert-butyl; and the variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty first, twenty second, twenty third, twenty fourth, and/or twenty fifth embodiments.
  • the invention provides a compound according to Structural Formula III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII(a), XIII(b), XIV, or a harmaceutically acceptable salt thereof, wherein R is
  • the present invention provides a compound represented by Structural Formula I'.
  • the invention provides a compound represented by Structural Formula I':
  • the thiazole ring is optionally substituted with -F or -CI;
  • X 4 is NR a or O
  • X 5 and X 6 are each independently NR b or O;
  • R 1 is (Ci-C 5 )alkyl
  • R is (Ci-C5)alkyl, -(C3-C 7 )cycloalkyl, or -(CH 2 ) q heterocyclyl (wherein the heterocycyl is a monocyclic 3-7-membered heterocyclic ring optionally substituted with one or more occurences of methyl), or benzyl (wherein the benzyl ring is optionally substituted with one or more occurences of halogen, methoxy, halomethoxy, methyl, halomethyl, or cyano);
  • each of R , R b , and R c is independently hydrogen or methyl
  • R d is independently halogen, methoxy, halomethoxy, methyl, halomethyl, or cyano; m is 0, 1, 2, or 3;
  • n 0, 1, or 2;
  • q is 0 or 1.
  • the invention provides a compound represented by
  • the invention provides a compound represented by Structural Formula I'-2:
  • the invention provides a compound represented by Structural Formula I'-3:
  • the invention provides a compound represented by Structural Formula I'-4:
  • the invention provides a compound according to Structural Formula I', I'-l, I'-2, 1'-3, or I'-4, or a pharmaceutically acceptable salt thereof, wherein X 4 is NH, and the remaining variables are as defined in the first embodiment.
  • the invention provides a compound according to Structural Formula I', I'-l, I'-2, 1'-3, or I'-4, or a pharmaceutically acceptable salt thereof, wherein R is (Q-G alkyl, -(C4-C6)cycloalkyl, -(CH 2 ) q heterocyclyl (wherein the heterocycyl is a monocyclic 4-6-membered heterocyclic ring optionally substituted with one methyl), or benzyl, and the remaining variables are as defined in the first and/or sixth embodiments.
  • R is isopropyl, ieri-butyl, cyclobutyl, cyclopentyl, oxetanyl,
  • R 3 is isopropyl or oxetanyl.
  • the invention provides a compound according to Structural Formula I', I'-l, I'-2, 1'-3, or I'-4, or a pharmaceutically acceptable salt thereof, wherein R d is halogen, and m is 0 or 1, and the remaining variables are as defined in the first, sixth,
  • the invention provides a compound according to Structural Formula I', I'-l, I'-2, 1'-3, or I'-4, or a pharmaceutically acceptable salt thereof, wherein R 1 is ie/t-butyl, and the remaining variables are as defined in the first, sixth, seventh, and/or eighth embodiments.
  • the invention provides a compound, or a pharmaceutically acceptable salt thereof wherein the compound is selected from the group consisting of:
  • the invention provides a compound represented by Structural Formula II':
  • the thiazole ring is optionally substituted with -F or -CI;
  • X 4 is NR a or O
  • X 5 and X 6 are each independently NR b or O;
  • R 1 is (Ci-C 5 )alkyl
  • R 4 is (Ci-C 4 )alkyl, -(C 3 -C 7 )cycloalkyl, -(CH(R c )) q -heterocycyl (wherein the heterocycyl is a monocyclic 3-7-membered heterocyclic ring optionally substituted with one or more occurences of methyl), -(CH(R c )) q -phenyl (wherein the phenyl ring is optionally substituted with one or more occurences of halogen, methoxy, halomethoxy, methyl, halomethyl, or cyano), or -(CH(R c )) q -2-pyridinyl (wherein the 2-pyridinyl ring is optionally substituted with one or more occurences of halogen, methoxy, halomethoxy, methyl, halomethyl, or cyano);
  • each of R , R b , and R c is independently hydrogen or methyl
  • n 0, 1, or 2;
  • the invention provides a compound represented by Structural Formula II'-l
  • the invention provides a compound represented by Structural Formula ⁇ -2:
  • the invention provides a compound according to Structural Formula II', II'-l or ⁇ -2, or a pharmaceutically acceptable salt thereof, wherein R 4 is isopropyl, oxetanyl, cyclobutyl, -CH 2 -2-pyrrolidinyl, -CH 2 -N-methyl-2-pyrroridinyl, - CH 2 -3-piperidinyl, -CH 2 -2-pyrazinyl, -CH 2 -2-pyrimidinyl, -CH(R C ) -phenyl, or -CH(R c )-2- pyridinyl, and that the phenyl and 2-pyridinyl rings are each independently and optionally
  • the invention provides a compound according to Structural Formula II', II'-l or ⁇ -2, or a pharmaceutically acceptable salt thereof, wherein X 4 is NH, and the remaining variables are as defined in the eleventh and/or fourteenth embodiments.
  • the invention provides a compound according to
  • the invention provides a compound, or a
  • pharmaceutically acceptable salt refers to a pharmaceutical salt that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, and allergic response, and is commensurate with a reasonable benefit/risk ratio.
  • Pharmaceutically- acceptable salts are well known in the art. For example, S. M. Berge et al. describes pharmacologically acceptable salts in J. Pharm. Sci., 1977, 66, 1-19.
  • Suitable pharmaceutically acceptable salts of the compounds disclosed herein include pharmaceutically acceptable salts with pharmaceutically acceptable acid(s).
  • Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include salts of inorganic acids (such as hydrochloric acid, hydrobromic, phosphoric, metaphosphoric, nitric, and sulfuric acids) and of organic acids (such as acetic acid, benzenesulfonic, benzoic, ethanesulfonic, methanesulfonic, succinic, and trifluoro acetic acid acids).
  • Compounds of the present teachings with acidic groups such as carboxylic acids can form pharmaceutically acceptable salts with pharmaceutically acceptable base(s).
  • Suitable pharmaceutically acceptable basic salts include ammonium salts, alkali metal salts (such as sodium and potassium salts) and alkaline earth metal salts (such as magnesium and calcium salts).
  • halo as used herein means halogen and includes fluoro, chloro, bromo and iodo.
  • alkyl used alone or as part of a larger moiety, such as “alkoxy” or
  • haloalkyl and the like, means saturated aliphatic straight-chain or branched monovalent hydrocarbon radical. Unless otherwise specified, an alkyl group typically has 1-5 carbon atoms, i.e. (Ci-C 5 )alkyl. As used herein, a "(Ci-C 5 )alkyl” group means a radical having from 1 to 5 carbon atoms in a linear or branched arrangement. Examples include methyl, ethyl, n- propyl, z ' so-propyl, and the like.
  • alkoxy means an alkyl radical attached through an oxygen linking atom, represented by -O-alkyl.
  • alkoxy means an alkyl radical attached through an oxygen linking atom, represented by -O-alkyl.
  • alkoxy includes methoxy, ethoxy, propoxy, and butoxy.
  • haloalkyl and “haloalkoxy” means alkyl or alkoxy, as the case may be, substituted with one or more halogen atoms.
  • alkylene group is a saturated aliphatic branched or straight-chain divalent hydrocarbon radical. Unless otherwise specified, an alkylene group typically has 2-6 carbon atoms, e.g. (C 2 -C 6 )alkylene.
  • alkenyl means branched or straight-chain monovalent hydrocarbon radical containing at least one double bond. Alkenyl may be mono or polyunsaturated, and may exist in the E or Z configuration. Unless otherwise specified, an alkenyl group typically has 2-6 carbon atoms, i.e., (C 2 -C 6 )alkenyl. For example, “(C 2 -C4)alkenyl” means a radical having from 2-4 carbon atoms in a linear or branched arrangement.
  • cycloalkyl means a monocyclic saturated hydrocarbon ring system.
  • a C3-C 6 cycloalkyl includes cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.
  • a "cycloalkyl” has from three to seven ring carbon atoms.
  • a bridged cycloalkyl means a bicyclic non-aromatic hydrocarbon ring system in which the two rings share at least three adjacent ring carbon atoms.
  • a bridged cycloalkyl typically has 6- 12 ring carbon atoms. Examples include, but are not limited to, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl, bicyclo[4.3.1]decyl, bicyclo[3.3.1]nonyl, bornyl, bornenyl, norbornyl, norbornenyl, 6,6- dimethylbicyclo [3.1.1]heptyl, tricyclo butyl, and adamantyl.
  • heterocyclyl means a saturated or unsaturated non-aromatic 4- 10 membered ring radical containing from 1 to 4 ring heteroatoms, which may be the same or different, selected from N, O, or S. It can be monocyclic, bicyclic or tricyclic (e.g. , a fused or bridged bicyclic or tricyclic ring).
  • Examples of include, but are not limited to, azetidinyl, morpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, dihydroimidazole, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, dihydropyrimidinyl, dihydrothienyl, dihydrothiophenyl, dihydrothiopyranyl, tetrahydroimidazole, tetrahydrofuranyl, tetrahydropyranyl,
  • a heterocyclic ring optionally contains one or more double bonds and/or is optionally fused with one or more aromatic rings (for example,
  • 3-7 membered monocyclic heterocyclic ring examples include, but are not limited to, azetidinyl, morpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, dihydroimidazole,
  • dihydrofuranyl dihydropyranyl, dihydropyridinyl, dihydropyrimidinyl, dihydrothienyl, dihydrothiophenyl, dihydrothiopyranyl, tetrahydroimidazole, tetrahydrofuranyl,
  • a bridged heterocyclyl means a bicyclic non-aromatic ring system containing from 1 to 4 ring heteroatoms in which the two rings share at least three adjacent ring atoms.
  • a bridged heterocyclyl typically has 6- 12 ring atoms.
  • Examples include, but are not limited to, azanorbornyl, quinuclidinyl, isoquinuclidinyl, tropanyl, azabicyclo[3.2.1]octanyl, azabicyclo[2.2.1]heptanyl, 2-azabicyclo[3.2.1]octanyl, azabicyclo[3.2.1]octanyl, azabicyclo[3.2.2]nonanyl, azabicyclo[3.3.0]nonanyl, and azabicyclo [3.3.1] no nanyl.
  • heteroaryl when used alone or as part of a larger moiety as in “heteroaralkyi” or
  • heteroarylalkoxy refers to aromatic ring groups having five to ten ring atoms selected from carbon and at least one (typically 1 to 4, more typically 1 or 2) heteroatoms (e.g. , oxygen, nitrogen or sulfur).
  • heteroaryl includes monocyclic rings and polycyclic rings in which a monocyclic heteroaromatic ring is fused to one or more other aromatic or
  • Heteroaryl includes monocyclic and bicyclic ring systems.
  • “Monocyclic 5-6 membered heteroaromatic ring (or heteroaryl)” means a monocyclic heteroaromatic ring having five or six ring atoms selected from carbon and at least one (typically 1 to 3, more typically 1 or 2) heteroatoms (e.g., oxygen, nitrogen or sulfur).
  • Examples of monocyclic 5-6 membered heteroaromatic ring groups include furanyl (e.g. , 2- furanyl, 3-furanyl), imidazolyl (e.g. , N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), isoxazolyl (e.g. , 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl), oxadiazolyl (e.g. , 2-oxadiazolyl, 5- oxadiazolyl), oxazolyl (e.g. , 2-oxazolyl, 4-oxazolyl, 5-oxazolyl), pyrazolyl (e.g.
  • pyrrolyl e.g. , 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl
  • pyridyl e.g. , 2-pyridyl, 3- pyridyl, 4-pyridyl
  • pyrimidinyl e.g. , 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl
  • pyridazinyl e.g. , 3-pyridazinyl
  • thiazolyl e.g. , 2-thiazolyl, 4-thiazolyl, 5-thiazolyl
  • isothiazolyl triazolyl (e.g. , 2-triazolyl, 5-triazolyl), tetrazolyl (e.g. , tetrazolyl), and thienyl (e.g., 2-thienyl, 3 -thienyl).
  • a non-hydrogen substituent replaces a hydrogen on a carbon or nitrogen of the substituent.
  • a substituted alkyl is an alkyl wherein at least one non-hydrogen substituent is in the place of a hydrogen substituent on the alkyl substituent.
  • monofluoroalkyl is alkyl substituted with a fluoro substituent
  • difluoroalkyl is alkyl substituted with two fluoro substituents. It should be recognized that if there is more than one substitution on a substituent, each non- hydrogen substituent can be identical or different (unless otherwise stated).
  • many moieties e.g.
  • alkyl, cycloalkyl, or a heterocyclic ring are referred to as being either "substituted” or “optionally substituted”.
  • a moiety is modified by one of these terms, unless otherwise noted, it denotes that any portion of the moiety that is known to one skilled in the art as being available for substitution can be substituted, which includes one or more substituents. If more than one substituent is present, then each substituent is independently selected. Such means for substitution are well-known in the art and/or taught by the instant disclosure.
  • the optional substituents can be any substituents that are suitable to attach to the moiety.
  • impermissible substituent patterns e.g., methyl substituted with 5 different groups, and the like.
  • Such impermissible substitution patterns are clearly recognized by a person of ordinary skill in the art.
  • a group is described as being optionally substituted by "one or more" substituents, it denotes that the group is optionally substituted by one, two, three, four, five or six substituents.
  • a group is optionally substituted by 1-3 substituents.
  • a group is optionally substituted by 1-2 substituents.
  • a group is optionally substituted by one substituent.
  • Suitable substituents are those which do not have a significant adverse effect on the ability of the compound to inhibit RAD51.
  • Stereoisomers are compounds that differ only in their spatial arrangement.
  • Stereoisomers include all diastereomeric, enantiomeric, and epimeric forms as well as racemates and mixtures thereof.
  • geometric isomer refers to cyclic compounds having at least two substituents, wherein the two substituents are both on the same side of the ring (cis) or wherein the substituents are each on opposite sides of the ring (trans).
  • a disclosed compound is named or depicted by structure without indicating stereochemistry, it is understood that the name or the structure encompasses one or more of the possible stereoisomers, or geometric isomers, or a mixture of the encompassed stereoisomers or geometric isomers.
  • geometric isomer When a geometric isomer is depicted by name or structure, it is to be understood that the named or depicted isomer exists to a greater degree than another isomer, that is that the geometric isomeric purity of the named or depicted geometric isomer is greater than 50%, such as at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by weight. Geometric isomeric purity is determined by dividing the weight of the named or depicted geometric isomer in the mixture by the total weight of all of the geometric isomers in the mixture.
  • Racemic mixture means 50% of one enantiomer and 50% of is corresponding enantiomer.
  • a compound with one chiral center is named or depicted without indicating the stereochemistry of the chiral center, it is understood that the name or structure encompasses both possible enantiomeric forms (e.g. , both enantiomerically-pure,
  • Enantiomeric and diastereomeric mixtures can be resolved into their component enantiomers or stereoisomers by well-known methods, such as chiral-phase gas
  • Enantiomers and diastereomers also can be obtained from diastereomerically- or
  • a compound When a compound is designated by a name or structure that indicates a single enantiomer, unless indicated otherwise, the compound is at least 60%, 70%, 80%, 90%, 99% or 99.9% optically pure (also referred to as "enantiomerically pure").
  • Optical purity is the weight in the mixture of the named or depicted enantiomer divided by the total weight in the mixture of both enantiomers.
  • stereochemistry of a disclosed compound is named or depicted by structure, and the named or depicted structure encompasses more than one stereoisomer (e.g. , as in a diastereomeric pair), it is to be understood that one of the encompassed stereoisomers or any mixture of the encompassed stereoisomers is included. It is to be further understood that the stereoisomeric purity of the named or depicted stereoisomers at least 60%, 70%, 80%, 90%, 99% or 99.9% by weight. The stereoisomeric purity in this case is determined by dividing the total weight in the mixture of the stereoisomers encompassed by the name or structure by the total weight in the mixture of all of the stereoisomers.
  • the compounds disclosed therein are RAD51 inhibitors.
  • the pharmaceutical composition of the present invention comprises one or more RAD51 inhibitors, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.
  • “Pharmaceutically acceptable carrier” and “pharmaceutically acceptable diluent” refer to a substance that aids the formulation and/or administration of an active agent to and/or absorption by a subject and can be included in the compositions of the present disclosure without causing a significant adverse toxicological effect on the subject.
  • Non- limiting examples of pharmaceutically acceptable carriers and/or diluents include water, NaCl, normal saline solutions, lactated Ringer's, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydro xymethycellulose, polyvinyl pyrrolidine, and colors, and the like.
  • Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with or interfere with the activity of the compounds provided herein.
  • auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with or interfere with the activity of the compounds provided herein.
  • auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with or interfere with the activity of the compounds provided herein.
  • auxiliary agents such
  • compositions of the present teachings optionally include one or more pharmaceutically acceptable carriers and/or diluents therefor, such as lactose, starch, cellulose and dextrose.
  • pharmaceutically acceptable carriers and/or diluents therefor such as lactose, starch, cellulose and dextrose.
  • Other excipients such as flavoring agents; sweeteners; and preservatives, such as methyl, ethyl, propyl and butyl parabens, can also be included. More complete listings of suitable excipients can be found in the Handbook of Pharmaceutical Excipients (5 th Ed., Pharmaceutical Press (2005)). A person skilled in the art would know how to prepare formulations suitable for various types of administration routes.
  • the present invention provides a method of treating a subject with a disease which can be ameliorated by inhibition of RAD51, by administering to the subject an effective amount of one or more disclosed compounds, or a pharmaceutically acceptable salt thereof, or the corresponding pharmaceutical composition.
  • Diseases which can be ameliorated by inhibition of RAD51 include treating cancer, autoimmune disease, immune deficiency, or neurodegenerative disease.
  • described herein is a method of treating cancer, autoimmune disease, immune deficiency, or neurodegenerative disease, the method comprising administering a therapeutically effective dose of a composition as described herein, e.g., a composition comprising a compound of the present invention, to a subject in need of treatment for cancer, autoimmune disease, immune deficiency, or neurodegenerative disease.
  • a composition as described herein e.g., a composition comprising a compound of the present invention
  • the subject can be a subject determined to have an increased level of DNA damage occurring in one or more cell types relative to a reference level.
  • DNA damage refers to breaks, nicks, and mutations of the DNA present in a cell.
  • the DNA damage can comprise one or more of single-strand breaks ⁇ e.g., "nicks"), double strand breaks (DSBs), and mutations.
  • the DNA damage can be one or more DSBs.
  • “mutation” refers to a change or difference in the genetic material of a cell as compared to a reference wildtype cell, e.g. a deletion, an insertion, a SNP, a gene rearrangement, and/or the introduction of an exogenous gene or sequence.
  • the subject can be determined to have an increased level of DNA damage if the subject is determined to have an increased level and/or activity of a DNA damage process or DNA editing enzyme.
  • DNA damage process refers to any activity or process in a cell which causes one or more types of DNA damage to occur.
  • an increased level of DNA damage can be an increased level of mutations, e.g., by determining the overall mutation status in all or a portion of the genome of a cell.
  • An overall mutation status at least 2% greater, e.g. 2% greater or more, 3% greater or more, 5% greater or more, 10% greater or more, or 20% greater or more than the overall mutation status in a reference cell can be indicative of an increased, elevated, and/or significant level of a DNA editing enzyme activity.
  • the level of hyper mutations can be determined.
  • the overall mutation status in the whole genome or a portion thereof can be determined using FISH, whole genome sequencing, high throughput sequencing, exome sequencing, hybridization, and/or PCR.
  • the activity of a DNA editing enzyme can be measured by determining the level of
  • the DNA editing enzyme is AID.
  • a level of mutation in specific target genes including IGH, BCL6, MYC, BCL1 1 A, CD93, PIM1 and/or PAX5 which is at least 2% greater, e.g.
  • 2% greater or more, 3% greater or more, 5% greater or more, 10% greater or more, or 20% greater or more than the level of mutation in IGH, BCL6, MYC, BCL1 1 A, CD93, PIM1 and/or PAX5 in a reference cell can be indicative of an increased, elevated, and/or significant level of AID activity.
  • an increased level of DNA damage can be an increased level of double strand breaks (DSBs).
  • the level of DSBs can be determined, by way of non- limiting example, by karyotyping, by ⁇ - ⁇ 2 ⁇ foci formation, and/or by using FISH analysis to detect DNA double strand breaks, e.g. DNA breakage detection fish (DBD-FISH) (Volpi and Bridger, BioTechniques, Vol. 45, No. 4, October 2008, pp. 385-409).
  • DBD-FISH DNA breakage detection fish
  • an increased level of DNA damage can be an increased level of single strand breaks.
  • the level of single-strand breaks in DNA can be determined, by way of non-limiting example, by COMET assays, FISH, or the use of single-strand break- specific probes. Detection of DNA breaks, both single and double -stranded is known in the art and described further, at, e.g. , Kumari et al. EXCLI Journal 2009 7:44-62 and Motalleb et al. Research Journal of Applied Sciences, Engineering and Technology. 2012 4: 1888- 1894; each of which is incorporated by reference herein in its entirety.
  • an increased level of activity of a DNA damage process can comprise an increased level and/or activity of a DNA editing enzyme.
  • the technology described herein is directed to treating cells having an active DNA editing enzyme with a compound of the present invention.
  • the technology described herein is directed to treating cells having an increased level and/or activity of a DNA editing enzyme with a compound of the present invention.
  • DNA editing enzyme refers to an enzyme which normally catalyzes the mutation, exchange or excision of DNA segments, particularly enzymes which can generate or promote the generation of point mutations, DNA single strand breaks, DNA double-strand breaks or protein-DNA adducts.
  • a DNA editing enzyme, as referred to herein is not necessarily site- specific in its action. Similarly, it is not necessarily cell specific.
  • the cell is a B cell expressing a detectable amount of such an enzyme.
  • Non- limiting examples of DNA editing enzymes include, but are not limited to Recombination Activating Gene 1 (RAGl ; NCBI Gene ID: 5896), Recombination Activating Gene 1 (RAG2; NCBI Gene ID: 5897), Sporulation- specific protein 11 (SPOl 1 ; NCBI Gene ID: 23626), APOBEC family members a Type 1 Topoisomerase; a Type 2 Topoisomerase; and/or AID.
  • the DNA editing enzyme can be AID.
  • the DNA editing enzyme can be a member of the APOBEC (apolipoprotein B mRNA editing enzyme, catalytic polypeptide -like) family.
  • APOBEC family refers to a family of cytidine deaminase enzymes having an N-terminal zinc-dependent cytidine deaminase catalytic domain comprising and a C-terminal pseudocatalytic domain.
  • Non-limiting examples of APOBEC family members include AID, APOBEC 1 (e.g. , NCBI Gene ID: 339), APOBEC2 (e.g. , NCBI Gene ID: 10930),
  • APOBEC3A e.g. , NCBI Gene ID: 200315
  • APOBEC3C e.g., NCBI Gene ID: 27350
  • APOBEC3E e.g. , NCBI Gene ID: 140564
  • APOBEC3F e.g. , NCBI Gene ID:200316
  • APOBEC3G e.g. , NCBI Gene ID: 60489
  • APOBEC3H e.g. , NCBI Gene ID: 164668
  • APOBEC4 e.g. , NCBI Gene ID: 403314
  • the DNA editing enzyme can be a Type 1 topoisomerase. In some embodiments, the DNA editing enzyme can be a Type 2 topoisomerase.
  • Topoisomerases generate breaks in DNA to help uncoil or relax the strand.
  • Type II topoisomerases hydrolyze ATP to generate DSB cuts, while Type I topoisomerases generate single- stranded breaks.
  • Non- limiting examples of Type II topoisomerases can include topoisomerase II (e.g., NCBI Gene ID: 7153 and 7155).
  • Non-limiting examples of Type I topoisomerases can include topoisomerase I (e.g. , NCBI Gene ID: 7150).
  • Embodiments of the technology described herein are based on the discovery that the compounds described herein can inhibit DNA repair mechanisms, e.g. , homologous repair.
  • Activation- induced cytidine deaminase (AID, or AICDA, also known as ARP2, CDA2 or HIGM2)
  • ARP2 Activation- induced cytidine deaminase
  • APOBEC catalytic polypeptide -like
  • a method of causing cell death comprising detecting increased expression of a DNA-editing enzyme (e.g. AID) in a cell and thereafter contacting the cell with a compound of the present invention; thereby resulting in cell death.
  • a method of causing cell death comprising increasing expression of a DNA-editing enzyme (e.g. AID) in a cell and thereafter contacting the cell with a compound of the present invention; thereby resulting in cell death.
  • a method of causing cell death comprising administering to a cell a therapeutically effective amount of a DNA editing enzyme (e.g. AID) and thereafter contacting the cell with a compound of the present invention; thereby resulting in cell death.
  • AID encoded by the AICDA gene (NCBI Gene ID: 57379), is required for proper B- cell function and is most prominently expressed in centroblast B -cells.
  • the protein is involved in somatic hypermutation, gene conversion, and class-switch recombination of immunoglobulin genes.
  • AID is normally expressed almost exclusively in antigen- activated germinal center B-cells, where it initiates immunoglobulin isotype class switching (Manis et al. 2002, Trends Immunol, 23, 31-39; Chaudhuri and Alt, Nat Rev Immunol, 2004, 4, 541- 552; Longerich et al., Curr Opin Immunol, 2006, 18, 164- 174; Chaudhuri et al., Adv
  • AID expression is regulated by CD40 ligand, B-cell receptor, IL4R, or Toll-like receptor stimulation (Crouch et al., J Exp Med 2007 204: 1145- 1156; Muramatsu et al., J Biol Chem 1999 274: 18470-6). After activation, AID is transiently upregulated, induces point mutations or DNA double strand breaks in a sequence nonspecific manner within immunoglobulin genes, and is then downregulated (Longerich et al., Curr Opin Immunol, 2006, 18, 164- 176; Chaudhuri et al., Adv Immunol 2007, 94, 157-214).
  • AID is active in only a tiny population of normal cells (antigen- activated B-cells) at any given time.
  • the genomic rearrangements and mutations controlled by AID lead to the development of antigen-recognition diversity, receptor editing and lymphoid effector function required for functional adaptive immunity (Mills, et al. Immunol Rev 2003 194:77-95).
  • Robbiani et al. has reported off-target activities of AID in B- cells, especially c-myc/IgH translocations
  • AID expression accelerates the rate of tumor development in Bcl6 transgenic mice (Pasqualucci et al., 2008, Nat. Genet. 40, 108- 112).
  • deregulated AID does not necessarily cause malignancy or translocation-associated cancer on its own in B cells (Muto et al., 2006, Proc. Natl. Acad. Sci. USA 103, 2752-2757; Okazaki et al., 2003, J. Exp. Med. 197, 1173-1181; Shen et al., 2008, Mol. Immunol. 45, 1883-1892).
  • AID is not required for the development of plasmacytosis or plasmacytoma in IL-6 transgenic or pristane-treated mice, respectively (Kovalchuk et al., 2007, J. Exp. Med. 204, 2989-3001; Ramiro et al., 2004, J. Exp. Med. 200, 1103-1110).
  • most human B cell lymphoma- associated translocations do not involve c-myc, and many do not involve Ig genes (Kuppers, 2005, Oncogene 20, 5580-5594).
  • AID chronic lymphocytic leukemia
  • CLL chronic lymphocytic leukemia
  • AID expression has been shown to be correlated with blast crisis B lineage leukemia and therapy resistance in myeloid leukemia and to be associated with generally poor prognosis in chronic B lymphocytic leukemia (Mao et al., Br J Dermatol 2001, 145: 117-122; Chaudhuri et al., Nature 2004, 430:992-8).
  • a method of treatment comprising; (a) selecting a subject having cells that express elevated levels of activation- induced cytidine deaminase (AID); and (b) administering a therapeutically effective amount of an inhibitor of double strand break repair (e.g. a compound of the present invention) to the subject; wherein an elevated level of AID is a level of AID that is higher than the level of AID in cells of the same type from a healthy individual.
  • an inhibitor of double strand break repair e.g. a compound of the present invention
  • the cells expressing elevated levels of AID are B cells.
  • the B cell expressing elevated levels of AID is a cancerous B cells or a B cell associated with autoimmune disease.
  • the subject can be a human subject.
  • Methods provided herein treat cancers and/or autoimmune disorders by inhibiting DNA double strand break repair. This inhibition proves lethal to cells expressing AID, as AID generates widespread genomic breaks, and the treatment with a double strand break repair inhibitor prevents the repair of these lesions which are being generated by the cell itself. This results in cell death in the subject which is specific to the cells expressing AID, e.g.
  • cancerous B cells and/or autoimmune cells Accordingly, as described herein, in one embodiment there is a provided a treatment paradigm that selectively induces self-destruction of certain diseased cells, while reducing the unintended side effects in healthy tissues.
  • an increased level and/or activity of a DNA editing enzyme can be an increased level of DNA editing enzyme mRNA.
  • mRNA levels can be assessed using, e.g. , biochemical and molecular biology techniques such as Northern blotting or other hybridization assays, nuclease protection assay, reverse transcription (quantitative RT-PCR) techniques, RNA-Seq, high throughput sequencing and the like. Such assays are well known to those in the art.
  • nuclear "run-on” (or "run-off) transcription assays are used (see e.g. Methods in Molecular Biology, Volume: 49 , Sep-27- 1995, Page Range: 229- 238).
  • Arrays can also be used; arrays, and methods of analyzing mRNA using such arrays have been described previously, e.g. in EP0834575, EP0834576, W096/31622, U.S. Pat. No. 5,837,832 or WO98/30883.
  • WO97/10365 provides methods for monitoring of expression levels of a multiplicity of genes using high density oligonucleotide arrays.
  • a subject can be determined to have an increased level of DNA damage occurring in one or more cell types relative to a reference level if the subject has been exposed to an agent that is known to cause such DNA damage.
  • agents can include a viral infection with a DNA integrating virus (e.g. adeno- associated virus, retrovirus, human T-lymphotropic virus, HIV- 1, oncovirus, hepatitis virus, hepatitis B virus); DNA damaging chemicals (e.g. acetaldehyde, polycyclic aromatic hydrocarbons, benzenes, nitrosamines, tobacco smoke, aflatoxin, and the like); DNA damaging chemotherapeutic agents (e.g. bleomycin, mitomycin, nitrogen mustards (e.g.
  • MNU N-Nitroso-N-methylurea
  • BCNU carmustine
  • CCNU lomustine
  • MeCCNU semustine
  • tetrazines e
  • Exposure to such agents can be the result of an accident, infection and/or environmental exposure or the result of a
  • the increased level of DNA damage can be occurring in a cell type affected by the cancer, autoimmune disease, and/or neurodegenerative disease.
  • the subject is determined to have an increased level of DNA damage occurring in a cell selected from the group consisting of: a cancer cell; an immune system cell; or a nervous system cell.
  • the DNA editing enzyme can be AID.
  • the level of AID can be the level of AID in a blood cell. In some embodiments, the level of AID can be the level of AID in a B cell.
  • an increased level of AID can be a detectable level of AID, e.g. , as described below herein.
  • the subject can be a human subject.
  • Methods provided herein treat cancers and/or autoimmune disorders by inhibiting DNA double strand break repair. This inhibition proves lethal to cells expressing AID, as AID generates widespread genomic breaks, and the treatment with a double strand break repair inhibitor prevents the repair of these lesions which are being generated by the cell itself. This results in cell death in the subject which is specific to the cells expressing AID, e.g.
  • cancerous B cells and/or autoimmune cells Accordingly, as described herein, in one embodiment there is a provided a treatment paradigm that selectively induces self-destruction of certain diseased cells, while reducing the unintended side effects in healthy tissues.
  • the cancer to be treated is a type with high expression of a DNA editing enzyme. In certain embodiments, the cancer to be treated is a B-cell neoplasm. Another embodiment is a method of treating a cancer by administering to the subject an effective amount of one or more disclosed compounds, or a pharmaceutically acceptable salt thereof, or the corresponding pharmaceutical composition. In one aspect, the cancer is selected from the group consisting of lymphoma, leukemia, and a plasma cell neoplasm. In another aspect, the cancer selected from the group consisting of carcinoma and sarcoma.
  • the cancer to be treated is a lymphoma.
  • Lymphomas which can be treated by the disclosed methods include Non-Hodgkin' s lymphoma; Burkitt's lymphoma; small lymphocytic lymphoma; lymphoplasmacytic lymphoma; MALT
  • lymphoma follicular lymphoma; diffuse large B-cell lymphoma; and T-cell lymphoma.
  • Lymphoma is a malignancy in the lymphatic cells of the immune system (e.g. B cells, T cells, or natural killer (NK) cells). Lymphomas often originate in the lymph nodes and present as solid tumors. They can metastasize to other organs such as the brain, bone, or skin. Extranodal sites are often located in the abdomen. Lymphomas are closely related to the lymphoid leukemia and in some cases a particular form of cancer is categorized as both a lymphoma and a leukemia.
  • Leukemias which can be treated by the disclosed methods include acute
  • ALL lymphoblastic leukemia
  • B-cell leukemia B-cell acute remission
  • lymphoblastic leukemia chronic lymphocytic leukemia (CLL); acute myelogenous leukemia (AML); chronic myelogenous leukemia (CML); and T-cell acute lymphoblastic leukemia (T- ALL).
  • CLL chronic lymphocytic leukemia
  • AML acute myelogenous leukemia
  • CML chronic myelogenous leukemia
  • T- ALL T-cell acute lymphoblastic leukemia
  • the cancer to be treated is B-cell neoplasms, B-cell leukemia, B-cell acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, Burkitt's leukemia, acute myelogenous leukemia and/or T-ALL.
  • B-cell leukemia B-cell leukemia
  • B-cell acute lymphoblastic leukemia chronic lymphocytic leukemia
  • chronic myelogenous leukemia chronic myelogenous leukemia
  • Burkitt's leukemia acute myelogenous leukemia and/or T-ALL.
  • B-cell lymphoma a cancer referred to as "B-cell lymphoma” or a “B- cell leukemia.”
  • the cancer to be treated is chronic lymphocytic leukemia (CLL) or chronic myelogenous leukemia (CML).
  • the cancer to be treated is a plasma cell neoplasm.
  • plasma cell neoplasms include multiple myeloma; plasma cell myeloma; plasma cell leukemia and plasmacytoma.
  • Carcinomas which can be treated by the disclosed methods include colon cancer; liver cancer; gastric cancer; intestinal cancer; esophageal cancer; breast cancer; ovarian cancer; head and neck cancer; lung cancer; and thyroid cancer.
  • Sarcomas which can be treated by the disclosed methods include soft tissue sarcoma and bone sarcoma.
  • any cancer characterized by high levels of DNA damage and/or DNA editing enzyme expression can be treated with a compound as described herein, e.g. a compound of the present invention.
  • a compound of the present invention e.g. a compound of the present invention.
  • sarcomas, epithelial cell cancer (carcinomas), colon cancer, gastric cancer, intestinal cancer, liver cancer, hepatocellular cancer, breast cancer, thyroid cancer, esophageal cancer, lung cancer, brain cancer, head and neck cancer, melanoma, renal cancer, prostate cancer, hemangioma, rhabdomyosarcoma, chondrosarcoma, osteosarcoma, fibrosarcoma and cholangiocarcinoma may be characterized by high levels of a DNA editing enzyme expression, e.g. AID.
  • the cancer to be treated is colon cancer, liver cancer, gastric cancer, intestinal cancer, breast cancer, lung cancer, thyroid cancer and/or cholangiocarcinoma.
  • Specific cancers that can be treated by the disclosed methods include cancer of the bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, gastrointestine, gum, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate, skin, stomach, testis, tongue, or uterus.
  • the cancer may specifically be of the following histological type, though it is not limited to these: neoplasm, malignant; carcinoma; carcinoma, undifferentiated; giant and spindle cell carcinoma; sarcomas; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma;
  • gastrinoma malignant
  • cholangiocarcinoma hepatocellular carcinoma
  • hepatocellular carcinoma and cholangiocarcinoma hepatocellular carcinoma and cholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposis coli; solid carcinoma; carcinoid tumor, malignant; branchiolo- alveolar
  • adenocarcinoma papillary adenocarcinoma; chromophobe carcinoma; acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma;
  • nonencapsulating sclerosing carcinoma adrenal cortical carcinoma; endometroid carcinoma; skin appendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma;
  • ceruminous adenocarcinoma mucoepidermoid carcinoma; cystadenocarcinoma; papillary cystadenocarcinoma; papillary serous cystadenocarcinoma; mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cell carcinoma; infiltrating duct carcinoma; medullary carcinoma; lobular carcinoma; inflammatory carcinoma; paget's disease, mammary; acinar cell carcinoma; adenosquamous carcinoma; adenocarcinoma w/squamous metaplasia; thymoma, malignant; ovarian stromal tumor, malignant; thecoma, malignant; granulosa cell tumor, malignant; androblastoma, malignant; Sertoli cell carcinoma; leydig cell tumor, malignant; lipid cell tumor, malignant; paraganglioma, malignant; extra-mammary paraganglio
  • mesenchymoma malignant; brenner tumor, malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma, malignant; dysgerminoma; embryonal carcinoma; teratoma, malignant; struma ovarii, malignant; choriocarcinoma; mesonephroma, malignant;
  • hemangio sarcoma hemangioendothelioma, malignant; Kaposi's sarcoma;
  • hemangiopericytoma malignant; lymphangio sarcoma; osteosarcoma; juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma, malignant; mesenchymal
  • chondrosarcoma giant cell tumor of bone; Ewing' s sarcoma; odontogenic tumor, malignant; ameloblastic odonto sarcoma; ameloblastoma, malignant; ameloblastic fibrosarcoma;
  • pinealoma malignant; chordoma; glioma, malignant; ependymoma; astrocytoma;
  • astrocytoma protoplasmic astrocytoma; fibrillary astrocytoma; astroblastoma; glioblastoma;
  • oligodendroglioma oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma; ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactory neurogenic tumor;
  • meningioma malignant
  • neurofibrosarcoma neurilemmoma
  • malignant granular cell tumor, malignant; malignant lymphoma
  • Hodgkin's disease hodgkin's
  • paragranuloma malignant lymphoma, small lymphocytic
  • malignant lymphoma large cell, diffuse; malignant lymphoma, follicular; mycosis fungoides; other specified non-Hodgkin's lymphomas;
  • malignant histiocytosis multiple myeloma; mast cell sarcoma; immunoproliferative small intestinal disease; leukemia; lymphoid leukemia; plasma cell leukemia; erythro leukemia; lymphosarcoma cell leukemia; myeloid leukemia; basophilic leukemia; eosinophilic leukemia; monocytic leukemia; mast cell leukemia; megakaryoblastic leukemia; myeloid sarcoma; and hairy cell leukemia.
  • the cancer is characterized by mutations in the mutS homologues ⁇ e.g., MSH2, MSH3, and MSH6), mutL homologues (e.g. MLH1), or mismatch repair endonuclease PMS2.
  • Mutations are changes in the genetic code. They include point mutations and frameshift mutations. In a point mutation, one nucleotide is swapped out for another. Therefore, the mutation occurs at a single point or location within the DNA strand.
  • Frameshift mutations are due to either insertions or deletions of nucleotides. This causes the entire DNA strand to elongate or to shrink in size. Thus, frameshift mutations may alter all of the codons that occur after the deletion or insertion.
  • the mutations referred to herein include, but are not limited to, insertions, deletions, duplications, inversions, or other recognized point mutations. It has now been found that RAD51 inhibitors are particularly effective in treating cancers with mutations in MSH (e.g. MSH6), MLH, or PMS2.
  • MutS Homo log 2 is a protein that in humans is encoded by the MSH2 gene, which is located on chromosome 2.
  • MSH2 is a tumor suppressor gene and more specifically a caretaker gene that codes for a DNA mismatch repair (MMR) protein, MSH2, which forms a heterodimer with MSH6 to make the human MutSa mismatch repair complex. It also dimerizes with MSH3 to form the MutSP DNA repair complex.
  • MSH2 is involved in many different forms of DNA repair, including transcription-coupled repair, homologous recombination, and base excision repair. Examples of the mutations in MSH2 include, but are not limited to, g.47630253_47630254del, g.47702411_47702421del,
  • g.47630484_47630485insG g.47693838_47693839del, g.47693862del, g.47693864del, g.47693873del, g.47693880dup, g.47693913del, g.47693924_47693925dup, g.47630493del, g.47697730_47706125del, g.(47693948_47698103)_(47710367_?)del,
  • g.47705437_47705438insA g.47635551_47635552del, g.47705440_47705441del, g.47705461del, g.47705490del, g.47705494del, g.47705495del, g.47635557_47635558del, g.47705505del, g.47705535dup, g.47705547del, g.47705560_47705561dup, g.47705561dup, g.47705562dup, g.47705588del, g.47705608_47705609del, g.47705618dup, g.47705627dup, g.47635571_47635601delins(217), g.(47705659_47707834)_(47710367_?)del,
  • g.47707878_47707884del g.47707883del, g.47707895_47707905del, g.47707897del, g.47707901_47707902del, g.47707905_47707906del, g.47707921del, g.47635583dup, g.47635583_47635584del, g.47707969_47707973del, g.47707996_47707997ins(l 15), g.47708009_47708010del, g.(47708011_47709917)_(47710367_?)del,
  • MutS Homo log 3 is a human homologue of the bacterial mismatch repair protein MutS that participates in the mismatch repair (MMR) system.
  • MSH3 typically forms the heterodimer MutSP with MSH2 in order to correct long insertion/deletion loops and base- base mispairs in micro satellites during DNA synthesis.
  • Deficient capacity for MMR is found in approximately 15% of colorectal cancers, and somatic mutations in the MSH3 gene can be found in nearly 50% of MMR-deficient colorectal cancers. Examples of the mutations in MSH3 include, but are not limited to, g.79970809del.
  • MSH6 encodes MutS homologue 6 (MSH6), a member of the Mutator S (MutS) family of proteins that are involved in DNA mismatch repair (MMR).
  • MMR DNA mismatch repair
  • the MSH6 protein forms a heterodimer with MutS homologue 2 (MSH2) in both human and yeast.
  • Human MSH2/6 recognizes single base-base mismatches and short insertion/deletion loops. Upon recognition of a mismatch, MSH2/6 complex binds and exchanges ADP for ATP, resulting in a conformational change to the complex that precedes base pair dissolution, base excision, and repair.
  • MSH6 mutations include frameshift and/or nonsense mutations and can result in nonfunctional MSH6 and loss of protein expression. Examples include a frameshift mutation at MSH6 amino acid residue 290 and a compounding missense T1189I.
  • Inactivating MSH6 mutations can be detected in cancers by routine diagnostics methods. These methods include, but are not limited to, obtaining cancer cells and other diagnostic indicators such as peripheral blood mononuclear cells (PBMCs), PBMC subpopulations, circulating blasts (CD34+ cells), circulating tumor cells and circulating exosomescancer cells by biopsy and blood tests and by obtaining lymphatic or other bodily fluids.
  • PBMCs peripheral blood mononuclear cells
  • CD34+ cells circulating blasts
  • tumor cells and circulating exosomescancer cells by biopsy and blood tests and by obtaining lymphatic or other bodily fluids.
  • RNA sequencing RNA-Seq
  • microarray quantitative PCR
  • NanoStringTM gene expression panels or MSH6 protein by immunohistochemistry, flow cytometry, immunocytochemistry or Western blot.
  • Methods for identifying inactivating MSH6 mutations are disclosed in Houlleberghs H, Goverde A, Lusseveld J, Dekker M, Bruno MJ, et al. (2017) Suspected Lynch syndrome associated MSH6 variants: A functional assay to determine their pathogenicity.
  • mutations in MSH6 include, but are not limited to,
  • MutL homo log 1, colon cancer, nonpolyposis type 2 (E. coli) is a protein that in humans is encoded by the MLHl gene located on Chromosome 3. It is a gene commonly associated with hereditary nonpolyposis colorectal cancer. Examples of the mutations in MSH6 include, but are not limited to,
  • g.37038149del g.37038149dup, g.37081690_37081691del, g.37081691_37081692del, g.37081706_37081708del, g.37081710_3708171 ldel, g.37035053_37035066del, g.37038154del, g.37038154_37038157del, g.37081738_37081739del, g.37081740del, g.37081753dup, g.37081757_3708176 ldup, g.37081782_37081783insAAGT,
  • g.37081787_37081793delinsATTT g.(37081786_37083758)_(37083823_37089009)del
  • g.(37081786_37083758)_(37089175_37090007)del g.37083759del
  • g.37083780dup g.37083781_37083784del
  • g.37083781_37083784delCTCA g.37083808_37083809del
  • g.37083816del g.37086069_37089606del
  • g.37050382_37050383delinsAT g.37050382_37050383delinsCT
  • g.37050390_37050396del g.37052950_37060990del
  • g.(37053591_37055922)_(37059091_37061800)del g.37035105del, g.37055928dup, g.37035106_37035116del, g.37055938del, g.37035108del, g.37055972_37055975del, g.37055976_37055979del, g.37035111del, g.37055990dup, g.37035114del, g.37035116del, g.37056036del, g.37056037dup, g.37058993_37059001del,
  • Human PMS2 related genes are located at bands 7pl2, 7pl3, 7ql 1, and 7q22. Exons 1 through 5 of these homologues share high degree of identity to human PMS2.
  • the product of this gene is involved in DNA mismatch repair.
  • the protein forms a heterodimer with MLHl and this complex interacts with MSH2 bound to mismatched bases. Defects in this gene are associated with hereditary nonpolyposis colorectal cancer, with Turcot syndrome, and are a cause of supratentorial primitive neuroectodermal tumors.
  • the present invention provides a method of treating patients with Lynch syndrome to reduce the likelihood of from developing or treating cancers derieved from Lynch syndrome, by administering to the subject an effective amount of one or more disclosed compounds, or a pharmaceutically acceptable salt thereof, or the corresponding pharmaceutical composition.
  • Lynch syndrome is a hereditary disorder caused by a mutation in a mismatch repair gene in which affected individuals have a higher than normal chance of developing colorectal cancer, endometrial cancer, and various other types of aggressive cancers, often at a young age - also called hereditary nonpolyposis colon cancer (HNPCC).
  • HNPCC hereditary nonpolyposis colon cancer
  • MMR mismatch repair
  • Those with Lynch syndrome carry up to an 85% risk of contracting colon cancer as well as a higher than average risk for endometrial cancer, stomach cancer, pancreatic cancer, kidney/ureter tract cancer, hepatobiliary tract cancer, gastric tract cancer, prostate cancer, ovarian cancer, gallbladder duct cancer, brain cancer, small intestine cancer, breast cancer, and skin cancer.
  • the method is a method of treating cancer derived from Lynch syndrome, selected from the group consisting of colon cancer, endometrial cancer, stomach cancer, pancreatic cancer, kidney/ureter tract cancer, hepatobiliary tract cancer, gastric tract cancer, prostate cancer, ovarian cancer, gallbladder duct cancer, brain cancer, small intestine cancer, breast cancer, and skin cancer.
  • Lynch syndrome selected from the group consisting of colon cancer, endometrial cancer, stomach cancer, pancreatic cancer, kidney/ureter tract cancer, hepatobiliary tract cancer, gastric tract cancer, prostate cancer, ovarian cancer, gallbladder duct cancer, brain cancer, small intestine cancer, breast cancer, and skin cancer.
  • the method is a method of treating autoimmune disease.
  • autoimmune diseases include lupus erythematosus; Wiskott-Aldrich syndrome; autoimmune lymphoproliferative syndrome; myasthenia gravis; rheumatoid arthritis (RA); lupus nephritis; multiple sclerosis; systemic lupus erythematosis; discoid lupus; subacute cutaneous lupus erythematosus; cutaneous lupus erythematosus including chilblain lupus erythematosus; chronic arthritis; Sjogren's syndrome; inflammatory chronic rhino sinusitis; colitis; celiac disease; inflammatory bowel disease; Barrett's esophagus; inflammatory gastritis; autoimmune nephritis; autoimmune vasculitis; autoimmune hepatitis; autoimmune carditis; autoimmune encephalitis; autoimmune diabetes
  • the method is a method of treating immune deficiency selected from the group consisting of Autoimmune Lymphoproliferative
  • APS Autoimmune polyglandular syndrome type 1
  • CEDS Caspase Eight Deficiency State
  • CCD Chronic Granulomatous Disease
  • CVID Common Variable Immunodeficiency
  • CTLA4 Congenital Neutropenia Syndromes
  • HIES hyper-immunoglobulin E syndrome
  • Hyper-IgM Hyper-Immunoglobulin M
  • LAD Leukocyte adhesion deficiency
  • LRBA LRBA deficiency
  • PI3 Kinase disease PLCG2-associated antibody deficiency and immune dysregulation
  • SCID severe combined immunodeficiency
  • WAT3 STAT3 gain-of-function disease
  • Warts Hypogammaglobulinemia, Infections, and Myelokathexis Syndrome
  • WIMS X-Linked Agammaglobulinemia
  • XLP X-Linked Lymphoproliferative Disease
  • XMEN Disease XMEN Disease
  • immune deficiency refers to a condition in which a portion or some portions of cell components constituting an immune system are defective or dysfunction, so that a normal immune mechanism is damaged.
  • immune deficiency means a condition under which: congenital immunity and/or acquired immunity are suppressed and/or decreased.
  • the immune -deficiency subject is an immunocompromised subject.
  • immune deficiencies can include AIDS, hypogammaglobulinemia, agammaglobulinemia, granulocyte deficiency, chronic granulomatous disease, asplenia, SCID, complement deficiency, and/or sickle cell anemia.
  • the method is a method of treating a neurodegenerative disorder selected from the group consisting of multiple sclerosis, Parkinson's disease (PD), Alzheimer's disease (AD), Dentatorubropallidoluysian atrophy (DRPLA), Huntington's Disease (HD), Spinocerebellar ataxia Type 1 (SCA1),
  • a neurodegenerative disorder selected from the group consisting of multiple sclerosis, Parkinson's disease (PD), Alzheimer's disease (AD), Dentatorubropallidoluysian atrophy (DRPLA), Huntington's Disease (HD), Spinocerebellar ataxia Type 1 (SCA1),
  • SCA6 Spinocerebellar ataxia 6
  • SCA7 Spinocerebellar ataxia Type 7
  • SCA8 Spinocerebellar ataxia Type 8
  • SCA12 Spinocerebellar ataxia Type 12
  • SCA17 Spinobulbar Muscular Ataxia/Kennedy Disease
  • SBMA Fargile X syndrome
  • FAAXE Fragile XE mental retardation
  • DM Myotonic dystrophy
  • a "subject” is a mammal, preferably a human, but can also be an animal in need of veterinary treatment, e.g. , companion animals (e.g. , dogs, cats, and the like), farm animals (e.g., cows, sheep, pigs, horses, and the like) and laboratory animals (e.g. , rats, mice, guinea pigs, and the like).
  • companion animals e.g. , dogs, cats, and the like
  • farm animals e.g., cows, sheep, pigs, horses, and the like
  • laboratory animals e.g. , rats, mice, guinea pigs, and the like.
  • the methods disclosed herein further comprise coadministering an effective amount of a DNA repair inhibitor, a DNA damage response (DDR) inhibitor, a DNA damaging agent or an immunomodulatory agent to the subject being treated for cancer, in addition to an effective amount of a disclosed RAD51 inhibitor.
  • DDR DNA damage response
  • DNA damaging agent or an immunomodulatory agent to the subject being treated for cancer, in addition to an effective amount of a disclosed RAD51 inhibitor.
  • DNA repair inhibitor refers to any agent that targets
  • DNA repair inhibitors include: RPA inhibitors, APE1 inhibitors, DNA ligase inhibitors, DNA polymerase inhibitors, Parp inhibitors etc.
  • DNA damage response inhibitor refers to any agent that targets components/processes involved in detecting DNA lesions, signaling the presence of DNA damage, and/or promote the repair of DNA damage.
  • DNA damage response inhibitors include checkpoint inhibitors, ATM and ATR inhibitiors, DNA-PK inhibitors, etc.
  • DNA damaging agent refers to any agent that directly or indirectly damages DNA for which homologous recombination could repair the damage.
  • the DNA damaging agents is selected from the group consisting of: exposure to a DNA damaging chemical; exposure to a chemotherapeutic agent; exposure to a radiochemotherapy, and exposure to ionizing or ultraviolet radiation.
  • Specific examples of DNA-damaging chemotherapeutic agents include alkylating agents, nitrosoureas, anti-metabolites, plant alkaloids, plant extracts and radioisotopes.
  • chemotherapeutic agents also include DNA-damaging drugs, for example, 5-fluorouracil (5-FU), capecitabine, S- l (Tegafur, 5-chloro-2,4-dihydroxypyridine and oxonic acid), 5-ethynyluracil, arabinosyl cytosine (ara-C), 5-azacytidine (5-AC), 2',2'-difluoro-2'-deoxycytidine (dFdC), purine antimetabolites (mercaptopurine, azathiopurine, thioguanine), gemcitabine hydrochlorine (Gemzar), pentostatin, allopurinol, 2- fluoro- arabinosyl- adenine (2F-ara-A), hydroxyurea, sulfur mustard (bischloroetyhylsulfide), mechlorethamine, melphalan, chlorambucil, cyclophosphamide, ifosfamide
  • nucleic acid damaging treatments include radiation e.g. , ultraviolet (UV), infrared (IR), or .alpha.-, .beta.-, or .gamma.- radiation, as well as environmental shock, e.g., hyperthermia.
  • UV ultraviolet
  • IR infrared
  • Immunomodulatory agent means an agent that modulates an immune response to an antigen but is not the antigen or derived from the antigen.
  • Modulate refers to inducing, enhancing, suppressing, directing, or redirecting an immune response.
  • agents include immuno stimulatory agents, such as adjuvants, that stimulate (or boost) an immune response to an antigen but is not an antigen or derived from an antigen.
  • immunomodulatory agents include, but are not limited to, Toll-like Receptor (TLR) agonists and Toll-like Receptor (TLR) antagonists.
  • TLR Toll-like Receptor
  • TLR Toll-like Receptor
  • TLR Toll-like Receptor
  • TLR Toll-like Receptor
  • TLR Toll-like Receptor
  • Such agents also include immunosuppressants.
  • the immunomodulatory agent is selected from the group consisting of immune checkpoint modulators, Toll-like receptor (TLR) agonists, cell-based therapies, cytokines and cancer vaccines.
  • the subject is determined to have an increased level and/or activity of a DNA damage process or DNA editing enzyme. In one aspect of this
  • the DNA editing enzyme is selected from the group consisting of activation induced cytidine deaminase (AID or AICDA), APOBEC2, APOBEC3A, APOBEC3C, APOBEC3D, APOBEC3F, APOBEC3G, APOBEC3H, APOBEC4, a Type 1 Topoisomerase, a Type 2 Topoisomerase, Recombination Activating Gene 1 (RAG 1), and Recombination Activating Gene 2 (RAG2).
  • blood cells obtained from the subject have been determined to have a detectable level of activation- induced cytidine deaminase (AID).
  • AID activation- induced cytidine deaminase
  • B cells obtained from the subject have been determined to have a detectable level of activation- induced cytidine deaminase (AID).
  • AID activation- induced cytidine deaminase
  • the detectable level of activation- induced cytidine deaminase is statistically significantly higher than the level of AID expressed in unactivated B- cells or normal non- immune cells from a healthy subject.
  • an "effective amount” to the subject will depend on the mode of administration, the type, and severity of the disease, and on the characteristics of the subject, such as general health, age, sex, body weight, and tolerance to drugs. The skilled artisan will be able to determine appropriate dosages depending on these and other factors.
  • an "effective amount" of any additional therapeutic agent(s) will depend on the type of drug used.
  • Suitable dosages are known for approved therapeutic agents and can be adjusted by the skilled artisan according to the condition of the subject, the type of condition(s) being treated and the amount of a compound of the invention being used by following, for example, dosages reported in the literature and recommended in the Physician 's Desk Reference (57th ed., 2003).
  • a therapeutically effective amount means an amount when administered to the subject which results in beneficial or desired results, including clinical results, e.g., inhibits, suppresses or reduces the symptoms of the condition being treated in the subject as compared to a control.
  • a therapeutically effective amount can be given in unit dosage form (e.g., 0.1 mg to about 50 g per day, alternatively from 1 mg to about 5 grams per day).
  • administer refers to methods that may be used to enable delivery of compositions to the desired site of biological action. These methods include, but are not limited to, intraarticular (in the joints), intravenous, intramuscular, intratumoral, intradermal, intraperitoneal, subcutaneous, orally, topically, intrathecally, inhalationally, transdermally, rectally, and the like.
  • Administration techniques that can be employed with the agents and methods described herein are found in e.g., Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.
  • the disclosed RAD51 inhibitors can be co-administered with other therapeutic agents.
  • co-administration means to encompass
  • administration of two or more therapeutic agents to a single subject and are intended to include treatment regimens in which the agents are administered by the same or different route of administration or at the same or different times.
  • the one or more compounds described herein will be co- administered with other agents. These terms encompass administration of two or more agents to the subject so that both agents and/or their metabolites are present in the subject at the same time. They include simultaneous administration in separate compositions, administration at different times in separate compositions, and/or administration in a composition in which both agents are present.
  • the compounds described herein and the other agent(s) are
  • the compounds described herein and the other agent(s) are admixed in the composition.
  • the particular mode of administration and the dosage regimen will be selected by the attending clinician, taking into account the particulars of the case (e.g., the subject, the disease, the disease state involved, the particular treatment). Treatment can involve daily or multi-daily or less than daily (such as weekly or monthly etc.) doses over a period of a few days to months, or even years. However, a person of ordinary skill in the art would immediately recognize appropriate and/or equivalent doses looking at dosages of approved compositions for treating a a RAD51 mediated disease using the disclosed RAD51 inhibitors for guidance.
  • the compounds or the corresponding pharmaceutical compositions taught herein can be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art.
  • the compounds of the present teachings may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump or transdermal administration and the pharmaceutical compositions formulated accordingly.
  • Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal and topical modes of administration. Parenteral administration can be by continuous infusion over a selected period of time.
  • the pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration.
  • the composition is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous, subcutaneous, intramuscular, oral, intranasal, or topical administration to human beings.
  • the pharmaceutical composition is formulated for intravenous administration.
  • a compound of the present teachings may be incorporated with excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
  • solutions of a compound of the present teachings can generally be prepared in water suitably mixed with a surfactant such as hydro xypropylcellulose.
  • Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
  • Example 9 Synthesis of isopropyl (3-(N-(tert-butyl)sulfamoyl)-4-(2-((lS,4r)-4-(((((S)- l-methylpyrrolidin-2-yl)methoxy)carbonyl)amino)cyclohexyl)thiazol-5- yl)phenyl)carbamate.
  • trans-4-hydroxycyclohexanecarboxylic acid (1 g, 6.94 mmol, 1 eq.) in MeCN (10 mL)
  • HBTU 2.89 g, 7.63 mmol, 1.1 eq.
  • TEA 2.11 g, 20.81 mmol, 2.90 mL, 3 eq.
  • NH 4 C1 742.07 mg, 13.87 mmol, 2 eq.
  • Example 22 Synthesis of trans-isopropyl N-[4-[5-[2-(tert-butylsulfamoyl)-4-(4 - pyridylmethylcarbamoylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate.
  • Example 25 Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3- (pyridin-3-ylmethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
  • Example 27 Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(3- fluorobenzyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
  • Example 29 Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3- (pyridin-2-ylmethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
  • Example 31 Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((3- fluoropyridin-2-yl)methyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
  • Example 33 Synthesis of trans-4-piperidylmethyl N-[3-(tert-butylsulfamoyl)-4-[2-[4- ( isopropoxycarbonylamino )cyclohexyl]thiazol-5-yl]phenyl]carbamate .
  • Example 34 Synthesis of isopropyl ((lR,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((R)- l-methylpyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2- yl)cyclohexyl)carbamate.
  • Example 35 Synthesis of isopropyl ((lS,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((S)-l- methylpyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2- yl)cyclohexyl)carbamate.
  • Example 36 Synthesis of isopropyl ((lR,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((R)- pyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
  • Example 37 Synthesis of isopropyl ((lS,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((S)- pyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
  • Example 46 Synthesis of isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[3-(isopropoxycarbonyl amino)azetidin-l-yl]thiazol-5-yl]phenyl]carbamate.

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Abstract

This application is directed to inhibitors of RAD51 represented by the following structural formula, and methods for their use, such as to treat cancer, autoimmune diseases, immune deficiencies, or neurodegenerative diseases.

Description

RAD51 INHIBITORS
CROSS-REFERENCES TO RELATED APPLICATIONS
This application claims the benefit of U.S. Provisional Application No. 62/556,763, filed on September 11, 2017; and U.S. Provisional Application No. 62/711,959, filed on July 30, 2018. The entire teachings of the aforementioned applications are incorporated herein by reference.
FIELD OF THE INVENTION
This application is directed to inhibitors of RAD51, and methods for their use, such as to treat conditions including cancers, autoimmune diseases, immune deficiencies, and neurodegenerative diseases.
BACKGROUND OF THE INVENTION
RAD51 is a eukaryote gene. The protein encoded by this gene is a member of the RAD51 protein family which assists in repair of DNA double strand breaks. RAD51 family members are homologous to the bacterial RecA, Archaeal RadA and yeast RAD51. The protein is highly conserved in most eukaryotes, from yeast to humans. In humans, RAD51 is a 339-amino acid protein that plays a major role in homologous recombination of DNA during double strand break (DSB) repair. RAD51 catalyzes strand transfer between a broken sequence and its undamaged homologue to allow re-synthesis of the damaged region (see homologous recombination models).
Studies have demonstrated a sensitization to certain DNA damaging therapies associated with cellular defects in proteins that promote HR DNA repair. This sensitization is particularly dramatic for DNA cross-linking chemotherapeutic drugs (30-100 times) and ionizing radiation (3-5 times) (Godthelp et al., "Mammalian Rad51C contributes to DNA cross-link resistance, sister chromatid cohesion and genomic stability," Nucleic Acids Res., 30:2172-2182, 2002; Tebbs et al., "Correction of chromosomal instability and sensitivity to diverse mutagens by a cloned cDNA of the XRCC3 DNA repair gene," Proc. Natl. Acad. Sci. USA, 92:6354-6358, 1995; Takata et al., "Chromosome instability and defective
recombinational repair in knockout mutants of the five Rad51 paralogs," Mol. Cell Biol., 21:2858-2866, 2001; Liu et al., "XRCC2 and XRCC3, new human Rad51-family members, promote chromosome stability and protect against DNA cross-links and other damages," Mol. Cell, 1:783-793, 1998). Several groups have recently demonstrated that HR can be partially inhibited in order to sensitize cells to DNA damaging therapies. Inhibition of XRCC3 (a RAD51 paralog protein), has been demonstrated using a synthetic peptide corresponding to another paralog protein. This peptide sensitized Chinese Hamster Ovary (CHO) cells to cisplatin and inhibited the formation of sub-nuclear RAD51 foci in response to DNA damage (Connell et al., Cancer Res., 64:3002-3005, 2004). Other researchers have inhibited the expression of the RAD51 protein itself (Russell et al., Cancer Res., 63:7377-7383, 2003; Hansen et al., Int. J. Cancer, 105:472-479, 2003; Ohnishi et al., Biochem. Biophys. Res. Commun., 245:319-324, 1998; Ito et al., J. Gene Med., 7(8): 1044- 1052, 2005; Collins et al., Nucleic Acids Res., 29: 1534-1538, 2001) or blocked its function by over-expressing a dominant negative BRC peptide fragment derived from BRCA2 (Chen et al., J. Biol. Chem., 274:32931-32935, 1999). In view of the connection between increased sensitivity to certain DNA damaging therapies and cellular defects in HR DNA repair-related proteins, there is a need for additional compounds that inhibit RAD51.
SUMMARY OF THE INVENTION
Applicant has now discovered novel compounds which are effective inhibitors of RAD51 (see Examples 1-75). The RAD51 inhibitors of the present invention inhibit homologous recombination by altering the nucleocytoplasmic distribution of RAD51 following DNA damage induction. The RAD51 inhibitors of the present invention reduce the repair of AID-induced DNA double strand breaks, leading to AID-dependent cytotoxicity in both normal and malignant B -lymphocytes. Certain of these RAD51 inhibitors have superior cell permeability as measured in Caco-2 cells (see Example 76). Among the RAD51 inhibitors with good cell permeability, several have superior metabolic stability (as measured by a liver microsome assay, see Example 77) and exposure, including oral exposure (see Example 79).
The present invention provides a compound represented by Structural Formula I:
Figure imgf000003_0001
I;
or a pharmaceutically acceptable salt thereof. The definition of each variable is provided below. The present invention also provides a pharmaceutical composition comprising a compound as described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or diluent.
The present invention further provides a method of treating a cancer, an autoimmune disease, an immune deficiency, or a neurodegenerative disease. The method comprises administering to a subject in need thereof an effective amount of a compound of disclosed herein or a pharmaceutically acceptable salt thereof or a pharmaceutical composition disclosed herein.
Also provided is the use of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical compositions disclosed herein for the preparation of a medicament for the treatment of a cancer, an autoimmune disease, an immune deficiency, or a neurodegenerative disease.
Also provided is a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein for use in treating a cancer, an autoimmune disease, an immune deficiency, or a neurodegenerative disease.
Although Applicant does not wish to be bound by any mechanism, it is believed that the compounds of the invention inhibit RAD51 by binding to MDC1 and causing reduced ability to form active complexes of RAD51.
DETAILED DESCRIPTION
In a first embodiment, the invention provides a compound represented by Structural Formula I:
Figure imgf000004_0001
I;
or a pharmaceutically acceptable salt thereof, wherein:
the thiazole ring is optionally substituted with -F or -CI;
Cy is -(C3-C7)cycloalkyl, bridged (C6-C12) cycloalkyl, or a 4-12 membered heterocyclic ring, each of which is optionally substituted with one or more groups selected from the group consisting of halogen, -OH, (Q-G alkyl, and
Figure imgf000004_0002
when X5 is connected with a nitrogen ring atom of Cy, X5 is absent;
when X5 is connected with a carbon ring atom of Cy, X5 is NR or O; X6 is NRa or O;
R1 is (Ci-C5)alkyl;
R3 is (Ci-C5)alkyl, -CH2-phenyl, -(C3-C7)cycloalkyl, -CH2-(C3-C7)cycloalkyl, -CH2-monocyclic 3-7 membered heterocyclic ring, or monocyclic 3-7 membered heterocyclic ring, wherein the (Ci-C5)alkyl, -(C3-C7)cycloalkyl, phenyl or monocyclic 3-7 membered heterocyclic ring represented by R 3 or in the group represented by R 3 is optionally substituted with one or more groups selected from the group consisting of halogen, -OH, (Ci-C4)alkyl, halomethyl, halomethoxy, -CN, and (Ci-C4)alkoxy;
R2 is -NRaC(0)0(Ci-C4)alkyl; -NRaC(0)NRa(Ci-C4)alkyl; -NRaC(0)0(C2- C4)alkenyl;
-NRaC(0)NRa(C2-C4)alkenyl; -NRaC(0)0-(C3-C6)cycloalkyl; -NRaC(0)NRa-(C3- C7)cycloalkyl; -NRaC(0)0-phenyl; -NRaC(0)NRa-phenyl; -NRaC(0)0-monocyclic 3-7 membered heterocyclic ring; -NR C(0)NR -monocyclic 3-7 membered heterocyclic ring; - NR C(0)0-monocyclic 5-6 membered heteroaromatic ring; -NR C(0)NR - monocyclic 5-6 membered heteroaromatic ring;
wherein the (Ci-C4)alkyl and the (C2-C4)alkenyl in the group represented by R are each optionally and independently substituted with one or more groups selected from the group consisting of halogen, N3, -OR , -NR R , -(C3-C6)cycloalkyl, phenyl, a monocyclic 3- 7 membered heterocyclic ring, and a monocyclic 5-6 membered heteroaromatic ring;
wherein the (C3-C7)cycloalkyl in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen, -CH3, =0, -OR and -NRaRa;
wherein the phenyl in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen, -CH3, halomethyl,
halomethoxy,
-CN, -ORa, and -N3;
wherein the heterocyclic ring in the group represented by R is optionally substituted with one or more groups selected from the group consisting of =0, halogen, -OR , -CH3, halomethyl, and halomethoxy;
wherein the heteroaromatic ring in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen, -CN, - CH3, halomethyl, halomethoxy, -ORa and -NRaRa; and
each R is independently -H or -CH3. In a second embodiment, the invention provides a compound represented by Structural Formula II:
Figure imgf000006_0001
or a pharmaceutically acceptable salt thereof, wherein
the thiazole ring is optionally substituted with -F or -CI;
Cy is cyclohexyl or a 6-membered monocyclic heterocyclic ring;
X5 and X6 are each independently NR or O;
R1 is (Ci-C5)alkyl;
R is (Ci-C5)alkyl or monocyclic 3-7-membered heterocyclic ring;
R2 is -NRaC(0)0(Ci-C4)alkyl; -NRaC(0)NRa(Ci-C4)alkyl; -NRaC(0)0(C2- C4)alkenyl;
-NRaC(0)NRa(C2-C4)alkenyl; -NRaC(0)-0(C3-C6)cycloalkyl; -NRaC(0)NRa-(C3- C6)cycloalkyl; -NRaC(0)0-phenyl; -NRaC(0)NRa-phenyl; -NRaC(0)0-monocyclic 3-7 membered heterocyclic ring; -NR C(0)NR -monocyclic 3-7 membered heterocyclic ring; - NR C(0)0-monocyclic 5-6 membered heteroaromatic ring; -NR C(0)NR - monocyclic 5-6 membered heteroaromatic ring;
wherein the (Ci-C4)alkyl and the (C2-C4)alkenyl in the group represented by R are each optionally and independently substituted with one or more halogen, N3, -OR , -NR R , -(C3-C6)cycloalkyl, phenyl, monocyclic 3-7-membered heterocyclic ring, or monocyclic 5-6-membered heteroaromatic ring;
wherein the -(C3-C6)cycloalkyl in the group represented by R is optionally substituted with one or more halogen, -CH3, -OR or -NR R ;
wherein the phenyl in the group represented by R is optionally substituted with one or more halogen, -CH3, halomethyl, halomethoxy, -OR , or -N3;
wherein the heterocyclic ring in the group represented by R is optionally substituted with one or more =0, halogen, -CH3, halomethyl, or halomethoxy;
wherein the heteroaromatic ring in the group represented by R is optionally substituted with one or more halogen, -CH3, halomethyl, halomethoxy, -OR or -NR R ; and each R is independently -H or -CH3. In a third embodiment, the invention provides a compound according to Structural Formula I, or a pharmaceutically acceptable salt thereof, wherein Cy is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl; azetidinyl, azepanyl, diazaspiro[4.4]nonyl, diazaspiro[3.5]nonyl, diazepanyl, dihydroimidazole, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, dihydropyrimidinyl, dihydrothienyl, dihydrothiophenyl,
dihydrothiopyranyl, hexahydropyridazinyl, hexahydropyrimidinyl, hydantoinyl, indolinyl, isoindolinyl, morpholinyl, oxiranyl, oxetanyl, piperidinyl, piperazinyl, pyrrolidinonyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydroimidazole, tetrahydroindolyl, tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridinyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, thio morpholinyl, tropanyl, valerolactamyl; bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl, bicyclo[4.3.1]decyl, bicyclo[3.3.1]nonyl, bornyl, bornenyl, norbornyl, norbornenyl, 6,6-dimethylbicyclo
[3.1.1]heptyl, tricyclobutyl, adamantly; azanorbornyl, quinuclidinyl, isoquinuclidinyl, tropanyl, azabicyclo[2.2.1]heptanyl, 2-azabicyclo[3.2.1]octanyl, azabicyclo[3.2.1]octanyl, azabicyclo[3.2.2]nonanyl, azabicyclo[3.3.0]nonanyl, azabicyclo [3.3.1] no nanyl,
diazabicyclo[2.2. l]heptanyl, diazabicyclo[3.2. l]octanyl, octahydropyrrolo[3,4-b]pyrrolyl, octahydropyrrolo[3,4-c]pyrrolyl; and the remaining variables are as defined in the first embodiment.
In a fourth embodiment, the invention provides a compound according to Structural Formula I or II, or a pharmaceutically acceptable salt thereof, wherein Cy is cyclohexyl, morpholinyl, thiomorpholinyl, piperidinyl, piperazinyl, hexahydropyridazinyl,
hexahydropyrimidinyl, valerolactamyl, dihydropyranyl, dihydropyridinyl,
dihydropyrimidinyl, dihydrothiopyranyl, tetrahydropyranyl, tetrahydropyridinyl,
tetrahydropyrimidinyl, or tetrahydrothiopyranyl; and the remaining variables are as defined in the first, second, and/or third embodiments.
In a fifth embodiment, the invention provides a compound represented by Structural Formula III,
Figure imgf000007_0001
or a pharmaceutically acceptable salt thereof, wherein:
X7 is NH or O; R4 is (Ci-C4)alkyl, (C3-C6)cycloalkyl, or a monocyclic 3-7 membered heterocyclic ring;
wherein the (Ci-C4)alkyl represented by R4 is optionally substituted with one or more groups selected from the group consisting of halogen, N3, -OR , -NR R ,
-(C3-C6)cycloalkyl, phenyl, a monocyclic 3-7 membered heterocyclic ring, and a monocyclic 5-6 membered heteroaromatic ring,
wherein the (C3-C6)cycloalkyl or the monocyclic 3-7 membered heterocyclic ring represented by R4, the (C3-C6)cycloalkyl or the monocyclic 3-7 membered heterocyclic ring in the group represented by R4 is optionally substituted with one or more groups selected from the group consisting of halogen, -OR , =0, and -CH3,
wherein the phenyl in the group represented by R4 is optionally substituted with one or more groups selected from the group consisting of halogen, -CH3, halomethyl,
halomethoxy,
-ORa, and -N3;
wherein the heteroaromatic ring in the group represented by R4 is optionally substituted with one or more groups selected from the group consisting of halogen and -CH3; and the remaining variables are as defined in the first, second, third, and/or fourth
embodiments.
In a sixth embodiment, the invention provides a compound according to Structural
Formula III, or a pharmaceutically acceptable salt thereof, wherein X 7 is NH or O; R 3 is (Ci-C5)alkyl; and R4 is (Q-G alkyl wherein the (Q-G alkyl represented by R4 is optionally substituted with one or more halogen, -OR , -NR R , -(C3-C6)cycloalkyl, phenyl (optionally substituted by one or more halogen, -CH3, halomethyl, halomethoxy, OR or N3), monocyclic 3-7-membered heterocyclic ring (optionally substituted by =0, halogen or - CH3), or monocyclic 5-6-membered heteroaromatic ring (optionally substituted by halogen or -CH3); and the remaining variables are as defined in the first, second, third, fourth and/or fifth embodiments.
In a seventh embodiment, the invention provides a compound represented by
Structural Formula IV,
Figure imgf000008_0001
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth and/or sixth embodiments.
In an eighth embodiment, the invention provides a compound represented by Structural Formula V,
Figure imgf000009_0001
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth and/or sixth embodiments.
In a ninth embodiment, the invention provides a compound represented by Structural Formula VI:
Figure imgf000009_0002
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth and/or sixth embodiments.
In a tenth embodiment, the invention provides a compound represented by Structural Formula VII:
Figure imgf000009_0003
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth and/or sixth embodiments.
In an eleventh embodiment, the invention provides a compound represented by
Structural Formula VIII:
Figure imgf000009_0004
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth and/or sixth embodiments. In a twelfth embodiment, the invention provides a compound represented by
Structural Formula IX:
Figure imgf000010_0001
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth and/or sixth embodiments.
In a thirteenth embodiment, the invention provides a compound according to
Structural Formula I, II, or III, or a pharmaceutically acceptable salt thereof, wherein Cy is azetidinyl or pyrrolidinyl, and the nitrogen ring atom is connected with the thiazole ring; and the remaining variables are as defined in the first, second, third, fourth, fifth and/or sixth embodiments.
In a fourteenth embodiment, the invention provides a compound according to
Structural Formula I, II, or III, or a pharmaceutically acceptable salt thereof, wherein Cy is l,7-diazaspiro[4.4]nonyl, 2,7-diazaspiro[4.4]nonyl, 2,7-diazaspiro[3.5]nonyl, 1,4-diazepanyl, 2,5-diazabicyclo[2.2. l]heptanyl, 3,8-diazabicyclo[3.2. l]octanyl, octahydropyrrolo[3,4- b]pyrrolyl, or octahydropyrrolo[3,4-c]pyrrolyl, and the two nitrogen ring atoms are connected with the thiazole ring and the -X5C(0)X6R3 moiety, respectively; and the remaining variables are as defined in the first, second, third, fourth, fifth and/or sixth embodiments.
In a fifteenth embodiment, the invention provides a compound according to Structural Formula III, IV, V, VI, VII, VIII, or IX, or a pharmaceutically acceptable salt thereof, wherein R4 is -(Ci-C3)alkyl, (C3-C6)cycloalkyl, or a monocyclic 3-7 membered heterocyclic ring, wherein the -(Ci-C3)alkyl is optionally substituted with (i) phenyl optionally substituted by one or more halogen or -CH3; (ii) a monocyclic 5-6 membered heteroaromatic ring optionally substituted by one or more halogen or -CH3; or (iii) a monocyclic 3-7 membered heterocyclic ring optionally substituted by one or more groups selected from the group consisting of halogen and -CH3; and the remaining variables are as defined in the first, second, third, fourth, fifth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, and/or fourteenth embodiments.
In a sixteenth embodiment, the invention provides a compound according to
Structural Formula III, IV, V, VI, VII, VIII, or IX, or a pharmaceutically acceptable salt thereof, wherein R4 is -(Ci-C3)alkyl, -CHR -phenyl, -CHR -5-6 membered heteraromatic ring, or -CHR -3-7 membered monocyclic heterocyclic ring, wherein the phenyl, 5-6 membered heteraromatic ring or 3-7 membered monocyclic heterocyclic ring in the group represented by R4 is optionally substituted one or more groups selected from the group consisting of halogen and -CH3; and the remaining variables are as defined in the first, second, third, fourth, fifth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, and/or fourteenth embodiments.
In a seventeenth embodiment, the invention provides a compound according to Structural Formula III, IV, V, VI, VII, VIII, or IX, or a pharmaceutically acceptable salt thereof, wherein R4 is -(Ci-C3)alkyl, optionally substituted with (i) phenyl optionally substituted by one or more halogen, -CH3, halomethyl, halomethoxy, OR , or N3; (ii) a monocyclic 5-6-membered heteroaromatic ring optionally substituted by one or more halogen or -CH3; or (iii) a monocyclic 3-7-membered heterocyclic ring optionally substituted by one or more =0 or -CH3; and the remaining variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, and/or fourteenth embodiments.
In an eighteenth embodiment, the invention provides a compound according to Structural Formula III, IV, V, VI, VII, VIII, or IX, or a pharmaceutically acceptable salt thereof, wherein R4 is -(Ci-C3)alkyl, optionally substituted with (i) phenyl optionally substituted by one or more halogen, -CH3, halomethyl, halomethoxy, OR , or N3; (ii) a monocyclic 5-6-membered heteroaromatic ring optionally substituted by one or more halogen or -CH3; or (iii) a monocyclic 3-7-membered heterocyclic ring optionally substituted by one or more =0 or -CH3; and the remaining variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, and/or seventeenth embodiments.
In a nineteenth embodiment, the invention provides a compound according to
Structural Formula I, III, IV, V, VI, VII, VIII, or IX, or a pharmaceutically acceptable salt thereof, wherein R is (Q-G alkyl, -(C4-C6)cycloalkyl, -CH2-phenyl, -CH2-monocyclic 4-6 membered heterocyclic ring, or monocyclic 4-6 membered heterocyclic ring, wherein the phenyl or monocyclic 4-6 membered heterocyclic ring represented by R or in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen,
-OR , and -CH3; and the remaining variables are as defined in the first, third, fourth, fifth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, and/or eighteenth embodiments.
In a twentieth embodiment, the invention provides a compound represented by Structural Formula X:
Figure imgf000012_0001
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenth embodiments.
In a twenty first embodiment, the invention provides a compound represented by Structural Formula XI:
Figure imgf000012_0002
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenth embodiments.
In a twenty second embodiment, the invention provides a compound represented by Structural Formula XII:
Figure imgf000012_0003
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth, sixth, eighth, fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenth embodiments. In a twenty third embodiment, the invention provides a compound represented by Structural Formula XIII(a) or XIII(b):
Figure imgf000013_0001
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth, sixth, eighth, fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenth embodiments.
In a twenty fourth embodiment, the invention provides a compound represented by Structural Formula XI
Figure imgf000013_0002
or a pharmaceutically acceptable salt thereof; and the variables are as defined in the first, second, third, fourth, fifth, sixth, tenth, fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenth embodiments.
In a twenty fifth embodiment, the invention provides a compound according to
Structural Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII(a), XIII(b), XIV, or a pharmaceutically acceptable salt thereof, wherein R is isopropyl, tert-buty\, cyclobutyl,
-N <**
cyclopentyl, benzyl, oxetanyl, tetrahydro-2H-pyranyl, or ; and the variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty first, twenty second, twenty third and/or twenty fourth embodiments. In an alternative embodiment, R 3 is isopropyl or oxetanyl. In another alternative embodiment, R 3 is isopropyl. In a twenty sixth embodiment, the invention provides a compound according to Structural Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII(a), XIII(b), XIV, or a pharmaceutically acceptable salt thereof, wherein R1 is tert-butyl; and the variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty first, twenty second, twenty third, twenty fourth, and/or twenty fifth embodiments.
In a twenty seventh embodiment, the invention provides a compound according to Structural Formula III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII(a), XIII(b), XIV, or a harmaceutically acceptable salt thereof, wherein R is
Figure imgf000014_0001
Figure imgf000014_0002
Figure imgf000014_0003
; and the variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth, nineteenth, twentieth, twenty first, twenty second, twenty third, twenty fourth, twenty fifth, and/or twenty sixth embodiments. In an
alternative embodiment, R
Figure imgf000014_0004
Figure imgf000014_0005
Figure imgf000015_0001
The present invention provides a compound represented by Structural Formula I'. In a first embodiment, the invention provides a compound represented by Structural Formula I':
Figure imgf000015_0002
or a pharmaceutically acceptable salt thereof, wherein:
the thiazole ring is optionally substituted with -F or -CI;
X4 is NRa or O;
X5 and X6 are each independently NRb or O;
R1 is (Ci-C5)alkyl;
R is (Ci-C5)alkyl, -(C3-C7)cycloalkyl, or -(CH2)qheterocyclyl (wherein the heterocycyl is a monocyclic 3-7-membered heterocyclic ring optionally substituted with one or more occurences of methyl), or benzyl (wherein the benzyl ring is optionally substituted with one or more occurences of halogen, methoxy, halomethoxy, methyl, halomethyl, or cyano);
each of R , Rb, and Rc is independently hydrogen or methyl;
Rd is independently halogen, methoxy, halomethoxy, methyl, halomethyl, or cyano; m is 0, 1, 2, or 3;
n is 0, 1, or 2; and
q is 0 or 1. In a second embodiment, the invention provides a compound represented by
Structural Formula I'-l:
Figure imgf000016_0001
or a pharmaceutically acceptable salt thereof, and the variables are as defined in the first embodiment.
In a third embodiment, the invention provides a compound represented by Structural Formula I'-2:
Figure imgf000016_0002
or a pharmaceutically acceptable salt thereof, and the variables are as defined in the first embodiment.
In a forth embodiment, the invention provides a compound represented by Structural Formula I'-3:
Figure imgf000016_0003
or a pharmaceutically acceptable salt thereof, and the variables are as defined in the first embodiment.
In a fifth embodiment, the invention provides a compound represented by Structural Formula I'-4:
Figure imgf000016_0004
or a pharmaceutically acceptable salt thereof, and the variables are as defined in the first embodiment. In a sixth embodiment, the invention provides a compound according to Structural Formula I', I'-l, I'-2, 1'-3, or I'-4, or a pharmaceutically acceptable salt thereof, wherein X4 is NH, and the remaining variables are as defined in the first embodiment.
In a seventh embodiment, the invention provides a compound according to Structural Formula I', I'-l, I'-2, 1'-3, or I'-4, or a pharmaceutically acceptable salt thereof, wherein R is (Q-G alkyl, -(C4-C6)cycloalkyl, -(CH2)qheterocyclyl (wherein the heterocycyl is a monocyclic 4-6-membered heterocyclic ring optionally substituted with one methyl), or benzyl, and the remaining variables are as defined in the first and/or sixth embodiments. In one specific embodiment, R is isopropyl, ieri-butyl, cyclobutyl, cyclopentyl, oxetanyl,
benzyl, tetrahydro-2H-pyranyl, or
Figure imgf000017_0001
In another specific embodiment, R3 is isopropyl or oxetanyl.
In an eighth embodiment, the invention provides a compound according to Structural Formula I', I'-l, I'-2, 1'-3, or I'-4, or a pharmaceutically acceptable salt thereof, wherein Rd is halogen, and m is 0 or 1, and the remaining variables are as defined in the first, sixth,
Figure imgf000017_0002
In a ninth embodiment, the invention provides a compound according to Structural Formula I', I'-l, I'-2, 1'-3, or I'-4, or a pharmaceutically acceptable salt thereof, wherein R1 is ie/t-butyl, and the remaining variables are as defined in the first, sixth, seventh, and/or eighth embodiments.
In a tenth embodiment, the invention provides a compound, or a pharmaceutically acceptable salt thereof wherein the compound is selected from the group consisting of:
Figure imgf000017_0003
Figure imgf000018_0001
In an eleventh embodiment, the invention provides a compound represented by Structural Formula II':
Figure imgf000018_0002
or a pharmaceutically acceptable salt thereof, wherein:
the thiazole ring is optionally substituted with -F or -CI;
X4 is NRa or O;
X5 and X6 are each independently NRb or O;
R1 is (Ci-C5)alkyl;
R4 is (Ci-C4)alkyl, -(C3-C7)cycloalkyl, -(CH(Rc))q-heterocycyl (wherein the heterocycyl is a monocyclic 3-7-membered heterocyclic ring optionally substituted with one or more occurences of methyl), -(CH(Rc))q-phenyl (wherein the phenyl ring is optionally substituted with one or more occurences of halogen, methoxy, halomethoxy, methyl, halomethyl, or cyano), or -(CH(Rc))q-2-pyridinyl (wherein the 2-pyridinyl ring is optionally substituted with one or more occurences of halogen, methoxy, halomethoxy, methyl, halomethyl, or cyano);
each of R , Rb, and Rc is independently hydrogen or methyl;
n is 0, 1, or 2; and
q is 0 or 1. In a twelveth embodiment, the invention provides a compound represented by Structural Formula II'-l
Figure imgf000019_0001
or a pharmaceutically acceptable salt thereof, and the variables are as defined in the eleventh embodiment.
In a thirteenth embodiment, the invention provides a compound represented by Structural Formula ΙΓ-2:
Figure imgf000019_0002
or a pharmaceutically acceptable salt thereof, and the variables are as defined in the eleventh embodiment.
In a fourteenth embodiment, the invention provides a compound according to Structural Formula II', II'-l or ΙΓ-2, or a pharmaceutically acceptable salt thereof, wherein R4 is isopropyl, oxetanyl, cyclobutyl, -CH2-2-pyrrolidinyl, -CH2-N-methyl-2-pyrroridinyl, - CH2-3-piperidinyl, -CH2-2-pyrazinyl, -CH2-2-pyrimidinyl, -CH(RC) -phenyl, or -CH(Rc)-2- pyridinyl, and that the phenyl and 2-pyridinyl rings are each independently and optionally
Figure imgf000019_0003
In a fifteenth embodiment, the invention provides a compound according to Structural Formula II', II'-l or ΙΓ-2, or a pharmaceutically acceptable salt thereof, wherein X4 is NH, and the remaining variables are as defined in the eleventh and/or fourteenth embodiments.
In a sixteenth embodiment, the invention provides a compound according to
Structural Formula II', II'-l or ΙΓ-2, or a pharmaceutically acceptable salt thereof, wherein R1 is tert-butyl, and the remaining variables are as defined in the eleventh, fourteenth, and fifteenth embodiments.
In a seventeenth embodiment, the invention provides a compound, or a
pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:
Figure imgf000020_0001
Also included are the compounds disclosed in the Exemplification, both in the pharmaceutically acceptable salt form and in the neutral form.
The term "pharmaceutically acceptable salt" refers to a pharmaceutical salt that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, and allergic response, and is commensurate with a reasonable benefit/risk ratio. Pharmaceutically- acceptable salts are well known in the art. For example, S. M. Berge et al. describes pharmacologically acceptable salts in J. Pharm. Sci., 1977, 66, 1-19.
Included in the present teachings are pharmaceutically acceptable salts of the compounds disclosed herein. Compounds having basic groups can form pharmaceutically acceptable salts with pharmaceutically acceptable acid(s). Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include salts of inorganic acids (such as hydrochloric acid, hydrobromic, phosphoric, metaphosphoric, nitric, and sulfuric acids) and of organic acids (such as acetic acid, benzenesulfonic, benzoic, ethanesulfonic, methanesulfonic, succinic, and trifluoro acetic acid acids). Compounds of the present teachings with acidic groups such as carboxylic acids can form pharmaceutically acceptable salts with pharmaceutically acceptable base(s). Suitable pharmaceutically acceptable basic salts include ammonium salts, alkali metal salts (such as sodium and potassium salts) and alkaline earth metal salts (such as magnesium and calcium salts).
Definitions
The term "halo" as used herein means halogen and includes fluoro, chloro, bromo and iodo.
The term "alkyl" used alone or as part of a larger moiety, such as "alkoxy" or
"haloalkyl" and the like, means saturated aliphatic straight-chain or branched monovalent hydrocarbon radical. Unless otherwise specified, an alkyl group typically has 1-5 carbon atoms, i.e. (Ci-C5)alkyl. As used herein, a "(Ci-C5)alkyl" group means a radical having from 1 to 5 carbon atoms in a linear or branched arrangement. Examples include methyl, ethyl, n- propyl, z'so-propyl, and the like.
The term "alkoxy" means an alkyl radical attached through an oxygen linking atom, represented by -O-alkyl. For example,
Figure imgf000021_0001
includes methoxy, ethoxy, propoxy, and butoxy.
The terms "haloalkyl" and "haloalkoxy" means alkyl or alkoxy, as the case may be, substituted with one or more halogen atoms.
An "alkylene group" is a saturated aliphatic branched or straight-chain divalent hydrocarbon radical. Unless otherwise specified, an alkylene group typically has 2-6 carbon atoms, e.g. (C2-C6)alkylene.
The term "alkenyl" means branched or straight-chain monovalent hydrocarbon radical containing at least one double bond. Alkenyl may be mono or polyunsaturated, and may exist in the E or Z configuration. Unless otherwise specified, an alkenyl group typically has 2-6 carbon atoms, i.e., (C2-C6)alkenyl. For example, "(C2-C4)alkenyl" means a radical having from 2-4 carbon atoms in a linear or branched arrangement.
The term "cycloalkyl" means a monocyclic saturated hydrocarbon ring system. For example, a C3-C6 cycloalkyl includes cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Unless otherwise described, a "cycloalkyl" has from three to seven ring carbon atoms.
A bridged cycloalkyl means a bicyclic non-aromatic hydrocarbon ring system in which the two rings share at least three adjacent ring carbon atoms. A bridged cycloalkyl typically has 6- 12 ring carbon atoms. Examples include, but are not limited to, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl, bicyclo[4.3.1]decyl, bicyclo[3.3.1]nonyl, bornyl, bornenyl, norbornyl, norbornenyl, 6,6- dimethylbicyclo [3.1.1]heptyl, tricyclo butyl, and adamantyl.
The terms "heterocyclyl", "heterocyclic ring", and "heterocyclic group", are used interchangeably herein, and means a saturated or unsaturated non-aromatic 4- 10 membered ring radical containing from 1 to 4 ring heteroatoms, which may be the same or different, selected from N, O, or S. It can be monocyclic, bicyclic or tricyclic (e.g. , a fused or bridged bicyclic or tricyclic ring). Examples of include, but are not limited to, azetidinyl, morpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, dihydroimidazole, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, dihydropyrimidinyl, dihydrothienyl, dihydrothiophenyl, dihydrothiopyranyl, tetrahydroimidazole, tetrahydrofuranyl, tetrahydropyranyl,
tetrahydrothienyl, tetrahydropyridinyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, and tetrahydrothiopyranyl. A heterocyclic ring optionally contains one or more double bonds and/or is optionally fused with one or more aromatic rings (for example,
tetrahydronaphthyridine, indolinone, dihydropyrrolotriazole, imidazopyrimidine,
quinolinone, dioxaspirodecane).
Examples of 3-7 membered monocyclic heterocyclic ring include, but are not limited to, azetidinyl, morpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, dihydroimidazole,
dihydrofuranyl, dihydropyranyl, dihydropyridinyl, dihydropyrimidinyl, dihydrothienyl, dihydrothiophenyl, dihydrothiopyranyl, tetrahydroimidazole, tetrahydrofuranyl,
tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridinyl, tetrahydropyrimidinyl,
tetrahydrothiophenyl, and tetrahydrothiopyranyl.
A bridged heterocyclyl means a bicyclic non-aromatic ring system containing from 1 to 4 ring heteroatoms in which the two rings share at least three adjacent ring atoms. A bridged heterocyclyl typically has 6- 12 ring atoms. Examples include, but are not limited to, azanorbornyl, quinuclidinyl, isoquinuclidinyl, tropanyl, azabicyclo[3.2.1]octanyl, azabicyclo[2.2.1]heptanyl, 2-azabicyclo[3.2.1]octanyl, azabicyclo[3.2.1]octanyl, azabicyclo[3.2.2]nonanyl, azabicyclo[3.3.0]nonanyl, and azabicyclo [3.3.1] no nanyl.
The terms "heteroaryl", "heteroaromatic", "heteroaryl ring", "heteroaryl group", "heteroaromatic ring", and "heteroaromatic group", are used interchangeably herein. "Heteroaryl" when used alone or as part of a larger moiety as in "heteroaralkyi" or
"heteroarylalkoxy", refers to aromatic ring groups having five to ten ring atoms selected from carbon and at least one (typically 1 to 4, more typically 1 or 2) heteroatoms (e.g. , oxygen, nitrogen or sulfur). "Heteroaryl" includes monocyclic rings and polycyclic rings in which a monocyclic heteroaromatic ring is fused to one or more other aromatic or
heteroaromatic rings. "Heteroaryl" includes monocyclic and bicyclic ring systems.
"Monocyclic 5-6 membered heteroaromatic ring (or heteroaryl)" means a monocyclic heteroaromatic ring having five or six ring atoms selected from carbon and at least one (typically 1 to 3, more typically 1 or 2) heteroatoms (e.g., oxygen, nitrogen or sulfur).
Examples of monocyclic 5-6 membered heteroaromatic ring groups include furanyl (e.g. , 2- furanyl, 3-furanyl), imidazolyl (e.g. , N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), isoxazolyl (e.g. , 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl), oxadiazolyl (e.g. , 2-oxadiazolyl, 5- oxadiazolyl), oxazolyl (e.g. , 2-oxazolyl, 4-oxazolyl, 5-oxazolyl), pyrazolyl (e.g. , 3-pyrazolyl, 4-pyrazolyl), pyrrolyl (e.g. , 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl), pyridyl (e.g. , 2-pyridyl, 3- pyridyl, 4-pyridyl), pyrimidinyl (e.g. , 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl), pyridazinyl (e.g. , 3-pyridazinyl), thiazolyl (e.g. , 2-thiazolyl, 4-thiazolyl, 5-thiazolyl), isothiazolyl, triazolyl (e.g. , 2-triazolyl, 5-triazolyl), tetrazolyl (e.g. , tetrazolyl), and thienyl (e.g., 2-thienyl, 3 -thienyl).
If a group is described as being "substituted," a non-hydrogen substituent replaces a hydrogen on a carbon or nitrogen of the substituent. Thus, for example, a substituted alkyl is an alkyl wherein at least one non-hydrogen substituent is in the place of a hydrogen substituent on the alkyl substituent. To illustrate, monofluoroalkyl is alkyl substituted with a fluoro substituent, and difluoroalkyl is alkyl substituted with two fluoro substituents. It should be recognized that if there is more than one substitution on a substituent, each non- hydrogen substituent can be identical or different (unless otherwise stated). As used herein, many moieties (e.g. , alkyl, cycloalkyl, or a heterocyclic ring) are referred to as being either "substituted" or "optionally substituted". When a moiety is modified by one of these terms, unless otherwise noted, it denotes that any portion of the moiety that is known to one skilled in the art as being available for substitution can be substituted, which includes one or more substituents. If more than one substituent is present, then each substituent is independently selected. Such means for substitution are well-known in the art and/or taught by the instant disclosure. The optional substituents can be any substituents that are suitable to attach to the moiety. A person of ordinary skill in the art will recognize that the compounds and definitions provided do not include impermissible substituent patterns (e.g., methyl substituted with 5 different groups, and the like). Such impermissible substitution patterns are clearly recognized by a person of ordinary skill in the art. When a group is described as being optionally substituted by "one or more" substituents, it denotes that the group is optionally substituted by one, two, three, four, five or six substituents. In one embodiment, a group is optionally substituted by 1-3 substituents. In one embodiment, a group is optionally substituted by 1-2 substituents. In one embodiment, a group is optionally substituted by one substituent.
Suitable substituents are those which do not have a significant adverse effect on the ability of the compound to inhibit RAD51. Where suitable substituents are not specifically enumerated, exemplary substituents include, but are not limited to, halo, -CN, alkyl, alkoxy, halomethyl, halomethoxy, (Ci-C5)alkyl, halo(Ci-C5)alkyl, (Ci-C5)alkoxy, -N02, -ORc , - NR Rb , -S(0)iRa', -NRaS(0)iRb', -S(0)iNRa Rb', -C(=0)ORa', -OC(=0)ORa', -C(=S)ORa', - 0(C=S)Ra',
-C(=0)NRa Rb', -NRa'C(=0)Rb', -C(=S)NRa Rb', -NRa C(=S)Rb', -NRa'(C=0)ORb',
-0(C=0)NRa Rb', -NRa'(C=S)ORb', -0(C=S)NRa Rb', -NR (C=0)NR Rb , - NR (C=S )NR Rb ,
-C(=S)R , -C(=0)R , (C3-C6)cycloalkyl, monocyclic heteroaryl and phenyl, wherein the (C3- C6)cycloalkyl, monocyclic heteroaryl and phenyl substituents are optionally and
independently substituted with -CH3, halomethyl, halo, methoxy or halomethoxy. Each R and each Rb are independently selected from -H and (Ci-C5)alkyl, wherein the (Ci-C5)alkyl group represented by R or Rb is optionally substituted with hydroxyl or (Ci-C3)alkoxy; Rc is -H, halo(Ci-C5)alkyl or (Ci-C5)alkyl, wherein the (Ci-C5)alkyl group represented by Rc is optionally substituted with hydroxyl or (Ci-C3)alkoxy; and i is 0, 1, or 2. =0 is also a suitable substituent for alkyl, cycloalkyl, and a heterocyclic ring.
Compounds having one or more chiral centers can exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement.
Stereoisomers include all diastereomeric, enantiomeric, and epimeric forms as well as racemates and mixtures thereof.
The term "geometric isomer" refers to cyclic compounds having at least two substituents, wherein the two substituents are both on the same side of the ring (cis) or wherein the substituents are each on opposite sides of the ring (trans). When a disclosed compound is named or depicted by structure without indicating stereochemistry, it is understood that the name or the structure encompasses one or more of the possible stereoisomers, or geometric isomers, or a mixture of the encompassed stereoisomers or geometric isomers.
When a geometric isomer is depicted by name or structure, it is to be understood that the named or depicted isomer exists to a greater degree than another isomer, that is that the geometric isomeric purity of the named or depicted geometric isomer is greater than 50%, such as at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by weight. Geometric isomeric purity is determined by dividing the weight of the named or depicted geometric isomer in the mixture by the total weight of all of the geometric isomers in the mixture.
Racemic mixture means 50% of one enantiomer and 50% of is corresponding enantiomer. When a compound with one chiral center is named or depicted without indicating the stereochemistry of the chiral center, it is understood that the name or structure encompasses both possible enantiomeric forms (e.g. , both enantiomerically-pure,
enantiomerically-enriched or racemic ) of the compound. When a compound with two or more chiral centers is named or depicted without indicating the stereochemistry of the chiral centers, it is understood that the name or structure encompasses all possible diastereomeric forms (e.g., diastereomerically pure, diastereomerically enriched and equimolar mixtures of one or more diastereomers (e.g., racemic mixtures) of the compound.
Enantiomeric and diastereomeric mixtures can be resolved into their component enantiomers or stereoisomers by well-known methods, such as chiral-phase gas
chromatography, chiral-phase high performance liquid chromatography, crystallizing the compound as a chiral salt complex, or crystallizing the compound in a chiral solvent.
Enantiomers and diastereomers also can be obtained from diastereomerically- or
enantiomerically-pure intermediates, reagents, and catalysts by well-known asymmetric synthetic methods.
When a compound is designated by a name or structure that indicates a single enantiomer, unless indicated otherwise, the compound is at least 60%, 70%, 80%, 90%, 99% or 99.9% optically pure (also referred to as "enantiomerically pure"). Optical purity is the weight in the mixture of the named or depicted enantiomer divided by the total weight in the mixture of both enantiomers.
When the stereochemistry of a disclosed compound is named or depicted by structure, and the named or depicted structure encompasses more than one stereoisomer (e.g. , as in a diastereomeric pair), it is to be understood that one of the encompassed stereoisomers or any mixture of the encompassed stereoisomers is included. It is to be further understood that the stereoisomeric purity of the named or depicted stereoisomers at least 60%, 70%, 80%, 90%, 99% or 99.9% by weight. The stereoisomeric purity in this case is determined by dividing the total weight in the mixture of the stereoisomers encompassed by the name or structure by the total weight in the mixture of all of the stereoisomers.
Pharmaceutical Compositions
The compounds disclosed therein are RAD51 inhibitors. The pharmaceutical composition of the present invention comprises one or more RAD51 inhibitors, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.
"Pharmaceutically acceptable carrier" and "pharmaceutically acceptable diluent" refer to a substance that aids the formulation and/or administration of an active agent to and/or absorption by a subject and can be included in the compositions of the present disclosure without causing a significant adverse toxicological effect on the subject. Non- limiting examples of pharmaceutically acceptable carriers and/or diluents include water, NaCl, normal saline solutions, lactated Ringer's, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydro xymethycellulose, polyvinyl pyrrolidine, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with or interfere with the activity of the compounds provided herein. One of ordinary skill in the art will recognize that other pharmaceutical excipients are suitable for use with disclosed compounds.
The pharmaceutical compositions of the present teachings optionally include one or more pharmaceutically acceptable carriers and/or diluents therefor, such as lactose, starch, cellulose and dextrose. Other excipients, such as flavoring agents; sweeteners; and preservatives, such as methyl, ethyl, propyl and butyl parabens, can also be included. More complete listings of suitable excipients can be found in the Handbook of Pharmaceutical Excipients (5th Ed., Pharmaceutical Press (2005)). A person skilled in the art would know how to prepare formulations suitable for various types of administration routes.
Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2003 - 20th edition) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19) published in 1999. The carriers, diluents and/or excipients are "acceptable" in the sense of being compatible with the other ingredients of the pharmaceutical composition and not deleterious to the recipient thereof.
Methods of Treatment
The present invention provides a method of treating a subject with a disease which can be ameliorated by inhibition of RAD51, by administering to the subject an effective amount of one or more disclosed compounds, or a pharmaceutically acceptable salt thereof, or the corresponding pharmaceutical composition. Diseases which can be ameliorated by inhibition of RAD51 include treating cancer, autoimmune disease, immune deficiency, or neurodegenerative disease.
In one aspect, described herein is a method of treating cancer, autoimmune disease, immune deficiency, or neurodegenerative disease, the method comprising administering a therapeutically effective dose of a composition as described herein, e.g., a composition comprising a compound of the present invention, to a subject in need of treatment for cancer, autoimmune disease, immune deficiency, or neurodegenerative disease.
In some embodiments, the subject can be a subject determined to have an increased level of DNA damage occurring in one or more cell types relative to a reference level. As used herein, "DNA damage" refers to breaks, nicks, and mutations of the DNA present in a cell. In some embodiments, the DNA damage can comprise one or more of single-strand breaks {e.g., "nicks"), double strand breaks (DSBs), and mutations. In some embodiments, the DNA damage can be one or more DSBs. As used herein, "mutation" refers to a change or difference in the genetic material of a cell as compared to a reference wildtype cell, e.g. a deletion, an insertion, a SNP, a gene rearrangement, and/or the introduction of an exogenous gene or sequence.
In some embodiments, the subject can be determined to have an increased level of DNA damage if the subject is determined to have an increased level and/or activity of a DNA damage process or DNA editing enzyme. As used herein, "DNA damage process" refers to any activity or process in a cell which causes one or more types of DNA damage to occur.
In some embodiments, an increased level of DNA damage can be an increased level of mutations, e.g., by determining the overall mutation status in all or a portion of the genome of a cell. An overall mutation status at least 2% greater, e.g. 2% greater or more, 3% greater or more, 5% greater or more, 10% greater or more, or 20% greater or more than the overall mutation status in a reference cell can be indicative of an increased, elevated, and/or significant level of a DNA editing enzyme activity. In some embodiments, the level of hyper mutations can be determined. In some embodiments, the overall mutation status in the whole genome or a portion thereof can be determined using FISH, whole genome sequencing, high throughput sequencing, exome sequencing, hybridization, and/or PCR. In some embodiments the activity of a DNA editing enzyme can be measured by determining the level of
hypermutations in the specific target genes including, but not limited to IGH, BCL6, MYC, BCL1 1A, CD93, PIM1 and/or PAX5. In certain embodiments the DNA editing enzyme is AID. In some embodiments, a level of mutation in specific target genes including IGH, BCL6, MYC, BCL1 1 A, CD93, PIM1 and/or PAX5 which is at least 2% greater, e.g. 2% greater or more, 3% greater or more, 5% greater or more, 10% greater or more, or 20% greater or more than the level of mutation in IGH, BCL6, MYC, BCL1 1 A, CD93, PIM1 and/or PAX5 in a reference cell can be indicative of an increased, elevated, and/or significant level of AID activity.
In some embodiments, an increased level of DNA damage can be an increased level of double strand breaks (DSBs). The level of DSBs can be determined, by way of non- limiting example, by karyotyping, by γ-Η2ΑΧ foci formation, and/or by using FISH analysis to detect DNA double strand breaks, e.g. DNA breakage detection fish (DBD-FISH) (Volpi and Bridger, BioTechniques, Vol. 45, No. 4, October 2008, pp. 385-409).
In some embodiments, an increased level of DNA damage can be an increased level of single strand breaks. The level of single-strand breaks in DNA can be determined, by way of non-limiting example, by COMET assays, FISH, or the use of single-strand break- specific probes. Detection of DNA breaks, both single and double -stranded is known in the art and described further, at, e.g. , Kumari et al. EXCLI Journal 2009 7:44-62 and Motalleb et al. Research Journal of Applied Sciences, Engineering and Technology. 2012 4: 1888- 1894; each of which is incorporated by reference herein in its entirety.
In some embodiments, an increased level of activity of a DNA damage process can comprise an increased level and/or activity of a DNA editing enzyme. In some embodiments, the technology described herein is directed to treating cells having an active DNA editing enzyme with a compound of the present invention. In some embodiments, the technology described herein is directed to treating cells having an increased level and/or activity of a DNA editing enzyme with a compound of the present invention. As used herein, "DNA editing enzyme" refers to an enzyme which normally catalyzes the mutation, exchange or excision of DNA segments, particularly enzymes which can generate or promote the generation of point mutations, DNA single strand breaks, DNA double-strand breaks or protein-DNA adducts. A DNA editing enzyme, as referred to herein, is not necessarily site- specific in its action. Similarly, it is not necessarily cell specific. In some embodiments, the cell is a B cell expressing a detectable amount of such an enzyme.
Non- limiting examples of DNA editing enzymes include, but are not limited to Recombination Activating Gene 1 (RAGl ; NCBI Gene ID: 5896), Recombination Activating Gene 1 (RAG2; NCBI Gene ID: 5897), Sporulation- specific protein 11 (SPOl 1 ; NCBI Gene ID: 23626), APOBEC family members a Type 1 Topoisomerase; a Type 2 Topoisomerase; and/or AID. In some embodiments, the DNA editing enzyme can be AID.
In some embodiments, the DNA editing enzyme can be a member of the APOBEC (apolipoprotein B mRNA editing enzyme, catalytic polypeptide -like) family. As used herein "APOBEC family" refers to a family of cytidine deaminase enzymes having an N-terminal zinc-dependent cytidine deaminase catalytic domain comprising and a C-terminal pseudocatalytic domain. Non-limiting examples of APOBEC family members include AID, APOBEC 1 (e.g. , NCBI Gene ID: 339), APOBEC2 (e.g. , NCBI Gene ID: 10930),
APOBEC3A (e.g. , NCBI Gene ID: 200315), APOBEC3C (e.g., NCBI Gene ID: 27350), APOBEC3E (e.g. , NCBI Gene ID: 140564), APOBEC3F (e.g. , NCBI Gene ID:200316), APOBEC3G (e.g. , NCBI Gene ID: 60489), APOBEC3H (e.g. , NCBI Gene ID: 164668), and APOBEC4(e.g. , NCBI Gene ID: 403314).
In some embodiments, the DNA editing enzyme can be a Type 1 topoisomerase. In some embodiments, the DNA editing enzyme can be a Type 2 topoisomerase.
Topoisomerases generate breaks in DNA to help uncoil or relax the strand. Type II topoisomerases hydrolyze ATP to generate DSB cuts, while Type I topoisomerases generate single- stranded breaks. Non- limiting examples of Type II topoisomerases can include topoisomerase II (e.g., NCBI Gene ID: 7153 and 7155). Non-limiting examples of Type I topoisomerases can include topoisomerase I (e.g. , NCBI Gene ID: 7150).
Embodiments of the technology described herein are based on the discovery that the compounds described herein can inhibit DNA repair mechanisms, e.g. , homologous repair. Activation- induced cytidine deaminase (AID, or AICDA, also known as ARP2, CDA2 or HIGM2), a DNA-editing enzyme that is a member of the apolipoprotein B mRNA editing enzymes, catalytic polypeptide -like (APOBEC), will cause widespread genomic breaks and cell death in cells with diminished homologous recombination ability (e.g. cells with diminished DNA double strand break repair abilities). Accordingly, provided herein is a method of causing cell death comprising detecting increased expression of a DNA-editing enzyme (e.g. AID) in a cell and thereafter contacting the cell with a compound of the present invention; thereby resulting in cell death. Accordingly, provided herein is a method of causing cell death comprising increasing expression of a DNA-editing enzyme (e.g. AID) in a cell and thereafter contacting the cell with a compound of the present invention; thereby resulting in cell death. Accordingly, provided herein is a method of causing cell death comprising administering to a cell a therapeutically effective amount of a DNA editing enzyme (e.g. AID) and thereafter contacting the cell with a compound of the present invention; thereby resulting in cell death.
AID, encoded by the AICDA gene (NCBI Gene ID: 57379), is required for proper B- cell function and is most prominently expressed in centroblast B -cells. The protein is involved in somatic hypermutation, gene conversion, and class-switch recombination of immunoglobulin genes. AID is normally expressed almost exclusively in antigen- activated germinal center B-cells, where it initiates immunoglobulin isotype class switching (Manis et al. 2002, Trends Immunol, 23, 31-39; Chaudhuri and Alt, Nat Rev Immunol, 2004, 4, 541- 552; Longerich et al., Curr Opin Immunol, 2006, 18, 164- 174; Chaudhuri et al., Adv
Immunol 2007, 94, 157-214). AID is required for somatic hypermutation and
immunoglobulin class switching in activated B cells. AID expression is regulated by CD40 ligand, B-cell receptor, IL4R, or Toll-like receptor stimulation (Crouch et al., J Exp Med 2007 204: 1145- 1156; Muramatsu et al., J Biol Chem 1999 274: 18470-6). After activation, AID is transiently upregulated, induces point mutations or DNA double strand breaks in a sequence nonspecific manner within immunoglobulin genes, and is then downregulated (Longerich et al., Curr Opin Immunol, 2006, 18, 164- 176; Chaudhuri et al., Adv Immunol 2007, 94, 157-214). Overall, AID is active in only a tiny population of normal cells (antigen- activated B-cells) at any given time. The genomic rearrangements and mutations controlled by AID lead to the development of antigen-recognition diversity, receptor editing and lymphoid effector function required for functional adaptive immunity (Mills, et al. Immunol Rev 2003 194:77-95). Recently it has been reported that AID has off-target point mutation activities (Liu, M. et al., Nature 2008, 451, 841-845; Liu and Schatz, Trends Immunol. 2009, 30, 173- 181 ; Perez-Duran et al., Carcinogenesis. 2007, 28(12):2427-33). Robbiani et al. has reported off-target activities of AID in B- cells, especially c-myc/IgH translocations
(Robbiani et al., Mol Cell 2009, 36(4):631-41). AID expression accelerates the rate of tumor development in Bcl6 transgenic mice (Pasqualucci et al., 2008, Nat. Genet. 40, 108- 112). However, deregulated AID does not necessarily cause malignancy or translocation-associated cancer on its own in B cells (Muto et al., 2006, Proc. Natl. Acad. Sci. USA 103, 2752-2757; Okazaki et al., 2003, J. Exp. Med. 197, 1173-1181; Shen et al., 2008, Mol. Immunol. 45, 1883-1892). In addition, despite its obligate role in c-myc/IgH translocation, AID is not required for the development of plasmacytosis or plasmacytoma in IL-6 transgenic or pristane-treated mice, respectively (Kovalchuk et al., 2007, J. Exp. Med. 204, 2989-3001; Ramiro et al., 2004, J. Exp. Med. 200, 1103-1110). However, most human B cell lymphoma- associated translocations do not involve c-myc, and many do not involve Ig genes (Kuppers, 2005, Oncogene 20, 5580-5594).
Overexpression of AID has been reported in chronic lymphocytic leukemia (CLL) (Hancer et al. Leuk Lymphoma. 2011 Jan;52(l):79-84; Heintel et al., Leukemia. 2004 Apr; 18(4):756-62). Further, AID expression has been shown to be correlated with blast crisis B lineage leukemia and therapy resistance in myeloid leukemia and to be associated with generally poor prognosis in chronic B lymphocytic leukemia (Mao et al., Br J Dermatol 2001, 145: 117-122; Chaudhuri et al., Nature 2004, 430:992-8). Further expression of AID in tumor cells from a variety of cancers has been reported including but not limited to lung, breast, gastric, colon, intestinal, liver cancer and choriangiocarcinoma (Greeve et al., Blood 2003, 1010, 3574-3580; Feldhahn et al., J Exp Med 2007, 204, 1157-1166; Kotani et al., PNAS USA 2007, 104, 1616-1620; Engels et al., 2008, Appl Immunohistochem Mol Morphol 16, 521-529; Klemm et al., 2009, Cancer Cell 6, 232-245; Palacios et al., 2010, Blood 115(22), 4488-4496; Leuenberger et al., 2009, Mod Pathol 32, 177-186; Gruber et al., 2010, Cancer Res 70, 7411-7420; inflammatory cancer (Marusawa 2008, Int J Biochem Cell Biol.40, 399- 402); follicular lymphoma (Hardianti et al., 2004, Leukemia 18, 826-831; Shikata et al., 2012, Cancer Sci. 103(3):415-21); thyroid cancer (Qiu et al. 2012, Mod Pathol 25(l),36-45); breast cancer (Borchert et al. 2011, BMC Cancer 11:347); Marusawa, et al., 2011, Adv Immunol 111 : 109-41; Zhang et al. 2012, Hum Pathol 43(3):423-34; Komori et al., 2008, Hepatology 47(3):888-896; Hockley 2010, Leukemia 24(5): 1084-6; adult T-cell leukemia (Nakamura et al., 2011, Br J Dermatol. 165(2):437-9). All of the references in the foregoing paragraph are incorporated by reference herein in their entireties.
Elevated levels of AID have been reported in arthritis (Xu et al. Scand. J. Immunol. 2009, 296, 2033-6) and in the MRL/Fas(lpr/lpr) mouse lupus model (White et al. 2011, Autoimmunity 44(8), 585-98). All of the references in the foregoing paragraph are
incorporated by reference herein in their entireties.
When DSB repair is inhibited, the extent of the DSBs generated by AID is much higher than previously suspected and the extent of genomic damage is so severe as to result in cell death. Accordingly, in one embodiment of the technology described herein, there is provided a method of treatment comprising; (a) selecting a subject having cells that express elevated levels of activation- induced cytidine deaminase (AID); and (b) administering a therapeutically effective amount of an inhibitor of double strand break repair (e.g. a compound of the present invention) to the subject; wherein an elevated level of AID is a level of AID that is higher than the level of AID in cells of the same type from a healthy individual. In some embodiments, the cells expressing elevated levels of AID are B cells. In some embodiments, the B cell expressing elevated levels of AID is a cancerous B cells or a B cell associated with autoimmune disease. In some embodiments, the subject can be a human subject.
Methods provided herein treat cancers and/or autoimmune disorders by inhibiting DNA double strand break repair. This inhibition proves lethal to cells expressing AID, as AID generates widespread genomic breaks, and the treatment with a double strand break repair inhibitor prevents the repair of these lesions which are being generated by the cell itself. This results in cell death in the subject which is specific to the cells expressing AID, e.g.
cancerous B cells and/or autoimmune cells. Accordingly, as described herein, in one embodiment there is a provided a treatment paradigm that selectively induces self-destruction of certain diseased cells, while reducing the unintended side effects in healthy tissues.
In some embodiments, an increased level and/or activity of a DNA editing enzyme can be an increased level of DNA editing enzyme mRNA. mRNA levels can be assessed using, e.g. , biochemical and molecular biology techniques such as Northern blotting or other hybridization assays, nuclease protection assay, reverse transcription (quantitative RT-PCR) techniques, RNA-Seq, high throughput sequencing and the like. Such assays are well known to those in the art. In one embodiment, nuclear "run-on" (or "run-off) transcription assays are used (see e.g. Methods in Molecular Biology, Volume: 49 , Sep-27- 1995, Page Range: 229- 238). Arrays can also be used; arrays, and methods of analyzing mRNA using such arrays have been described previously, e.g. in EP0834575, EP0834576, W096/31622, U.S. Pat. No. 5,837,832 or WO98/30883. WO97/10365 provides methods for monitoring of expression levels of a multiplicity of genes using high density oligonucleotide arrays.
In some embodiments, a subject can be determined to have an increased level of DNA damage occurring in one or more cell types relative to a reference level if the subject has been exposed to an agent that is known to cause such DNA damage. Non-limiting examples of such agents can include a viral infection with a DNA integrating virus (e.g. adeno- associated virus, retrovirus, human T-lymphotropic virus, HIV- 1, oncovirus, hepatitis virus, hepatitis B virus); DNA damaging chemicals (e.g. acetaldehyde, polycyclic aromatic hydrocarbons, benzenes, nitrosamines, tobacco smoke, aflatoxin, and the like); DNA damaging chemotherapeutic agents (e.g. bleomycin, mitomycin, nitrogen mustards (e.g.
mechlorethamine, cyclophosphamide, melphalan, chlorambucil, ifosfamide and busulfan), nitrosoureas (e.g., N-Nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU) and semustine (MeCCNU), fotemustine and streptozotocin), tetrazines (e.g. , dacarbazine, mitozolomide and temozolomide),aziridines (e.g. , thiotepa, mytomycin and diaziquone (AZQ)), cisp latins (e.g., cisp latin, carboplatin and oxalip latin) procarbazine and
hexamethylmelamine); and ionizing or ultraviolet radiation. Exposure to such agents can be the result of an accident, infection and/or environmental exposure or the result of a
therapeutic administration of such agents.
In some embodiments, the increased level of DNA damage can be occurring in a cell type affected by the cancer, autoimmune disease, and/or neurodegenerative disease. In some embodiments, the subject is determined to have an increased level of DNA damage occurring in a cell selected from the group consisting of: a cancer cell; an immune system cell; or a nervous system cell.
In some embodiments, the DNA editing enzyme can be AID. In some embodiments, the level of AID can be the level of AID in a blood cell. In some embodiments, the level of AID can be the level of AID in a B cell.
In some embodiments, an increased level of AID can be a detectable level of AID, e.g. , as described below herein.
In some embodiments, the subject can be a human subject.
Methods provided herein treat cancers and/or autoimmune disorders by inhibiting DNA double strand break repair. This inhibition proves lethal to cells expressing AID, as AID generates widespread genomic breaks, and the treatment with a double strand break repair inhibitor prevents the repair of these lesions which are being generated by the cell itself. This results in cell death in the subject which is specific to the cells expressing AID, e.g.
cancerous B cells and/or autoimmune cells. Accordingly, as described herein, in one embodiment there is a provided a treatment paradigm that selectively induces self-destruction of certain diseased cells, while reducing the unintended side effects in healthy tissues.
Methods of defecting cancers in patients with increased levels of DNA damage or increased levels of DNA editing enzymes are disclosed in WO2016/094897, incorporated herein by reference.
In certain embodiments, the cancer to be treated is a type with high expression of a DNA editing enzyme. In certain embodiments, the cancer to be treated is a B-cell neoplasm. Another embodiment is a method of treating a cancer by administering to the subject an effective amount of one or more disclosed compounds, or a pharmaceutically acceptable salt thereof, or the corresponding pharmaceutical composition. In one aspect, the cancer is selected from the group consisting of lymphoma, leukemia, and a plasma cell neoplasm. In another aspect, the cancer selected from the group consisting of carcinoma and sarcoma.
In certain embodiments, the cancer to be treated is a lymphoma. Lymphomas which can be treated by the disclosed methods include Non-Hodgkin' s lymphoma; Burkitt's lymphoma; small lymphocytic lymphoma; lymphoplasmacytic lymphoma; MALT
lymphoma; follicular lymphoma; diffuse large B-cell lymphoma; and T-cell lymphoma.
Lymphoma is a malignancy in the lymphatic cells of the immune system (e.g. B cells, T cells, or natural killer (NK) cells). Lymphomas often originate in the lymph nodes and present as solid tumors. They can metastasize to other organs such as the brain, bone, or skin. Extranodal sites are often located in the abdomen. Lymphomas are closely related to the lymphoid leukemia and in some cases a particular form of cancer is categorized as both a lymphoma and a leukemia.
Leukemias which can be treated by the disclosed methods include acute
lymphoblastic leukemia (ALL); Burkitt's leukemia; B-cell leukemia; B-cell acute
lymphoblastic leukemia; chronic lymphocytic leukemia (CLL); acute myelogenous leukemia (AML); chronic myelogenous leukemia (CML); and T-cell acute lymphoblastic leukemia (T- ALL).
In certain embodiments the cancer to be treated is B-cell neoplasms, B-cell leukemia, B-cell acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, Burkitt's leukemia, acute myelogenous leukemia and/or T-ALL. The maturation of B cells most typically ceases or substantially decreases when the foreign antigen has been neutralized. Occasionally, however, proliferation of a particular B cell will continue unabated; such proliferation can result in a cancer referred to as "B-cell lymphoma" or a "B- cell leukemia." In certain embodiments the cancer to be treated is chronic lymphocytic leukemia (CLL) or chronic myelogenous leukemia (CML).
In certain embodiments the cancer to be treated is a plasma cell neoplasm. Examples for plasma cell neoplasms include multiple myeloma; plasma cell myeloma; plasma cell leukemia and plasmacytoma.
Carcinomas which can be treated by the disclosed methods include colon cancer; liver cancer; gastric cancer; intestinal cancer; esophageal cancer; breast cancer; ovarian cancer; head and neck cancer; lung cancer; and thyroid cancer. Sarcomas which can be treated by the disclosed methods include soft tissue sarcoma and bone sarcoma.
Any cancer characterized by high levels of DNA damage and/or DNA editing enzyme expression can be treated with a compound as described herein, e.g. a compound of the present invention. For example, sarcomas, epithelial cell cancer (carcinomas), colon cancer, gastric cancer, intestinal cancer, liver cancer, hepatocellular cancer, breast cancer, thyroid cancer, esophageal cancer, lung cancer, brain cancer, head and neck cancer, melanoma, renal cancer, prostate cancer, hemangioma, rhabdomyosarcoma, chondrosarcoma, osteosarcoma, fibrosarcoma and cholangiocarcinoma may be characterized by high levels of a DNA editing enzyme expression, e.g. AID. In certain embodiments the cancer to be treated is colon cancer, liver cancer, gastric cancer, intestinal cancer, breast cancer, lung cancer, thyroid cancer and/or cholangiocarcinoma.
Specific cancers that can be treated by the disclosed methods include cancer of the bladder, blood, bone, bone marrow, brain, breast, colon, esophagus, gastrointestine, gum, head, kidney, liver, lung, nasopharynx, neck, ovary, prostate, skin, stomach, testis, tongue, or uterus. In addition, the cancer may specifically be of the following histological type, though it is not limited to these: neoplasm, malignant; carcinoma; carcinoma, undifferentiated; giant and spindle cell carcinoma; sarcomas; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma;
gastrinoma, malignant; cholangiocarcinoma; hepatocellular carcinoma; combined
hepatocellular carcinoma and cholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposis coli; solid carcinoma; carcinoid tumor, malignant; branchiolo- alveolar
adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma;
nonencapsulating sclerosing carcinoma; adrenal cortical carcinoma; endometroid carcinoma; skin appendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma;
ceruminous adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma; papillary cystadenocarcinoma; papillary serous cystadenocarcinoma; mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cell carcinoma; infiltrating duct carcinoma; medullary carcinoma; lobular carcinoma; inflammatory carcinoma; paget's disease, mammary; acinar cell carcinoma; adenosquamous carcinoma; adenocarcinoma w/squamous metaplasia; thymoma, malignant; ovarian stromal tumor, malignant; thecoma, malignant; granulosa cell tumor, malignant; androblastoma, malignant; Sertoli cell carcinoma; leydig cell tumor, malignant; lipid cell tumor, malignant; paraganglioma, malignant; extra-mammary paraganglioma, malignant; pheochromocytoma; glomangiosarcoma; malignant melanoma; amelanotic melanoma; superficial spreading melanoma; malig melanoma in giant pigmented nevus; epithelioid cell melanoma; blue nevus, malignant; sarcoma; fibrosarcoma; fibrous histiocytoma, malignant; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma; embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma; mixed tumor, malignant; mullerian mixed tumor; nephroblastoma; hepatoblastoma; carcinosarcoma;
mesenchymoma, malignant; brenner tumor, malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma, malignant; dysgerminoma; embryonal carcinoma; teratoma, malignant; struma ovarii, malignant; choriocarcinoma; mesonephroma, malignant;
hemangio sarcoma; hemangioendothelioma, malignant; Kaposi's sarcoma;
hemangiopericytoma, malignant; lymphangio sarcoma; osteosarcoma; juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma, malignant; mesenchymal
chondrosarcoma; giant cell tumor of bone; Ewing' s sarcoma; odontogenic tumor, malignant; ameloblastic odonto sarcoma; ameloblastoma, malignant; ameloblastic fibrosarcoma;
pinealoma, malignant; chordoma; glioma, malignant; ependymoma; astrocytoma;
protoplasmic astrocytoma; fibrillary astrocytoma; astroblastoma; glioblastoma;
oligodendroglioma; oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma; ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactory neurogenic tumor;
meningioma, malignant; neurofibrosarcoma; neurilemmoma, malignant; granular cell tumor, malignant; malignant lymphoma; Hodgkin's disease; hodgkin's; paragranuloma; malignant lymphoma, small lymphocytic; malignant lymphoma, large cell, diffuse; malignant lymphoma, follicular; mycosis fungoides; other specified non-Hodgkin's lymphomas;
malignant histiocytosis; multiple myeloma; mast cell sarcoma; immunoproliferative small intestinal disease; leukemia; lymphoid leukemia; plasma cell leukemia; erythro leukemia; lymphosarcoma cell leukemia; myeloid leukemia; basophilic leukemia; eosinophilic leukemia; monocytic leukemia; mast cell leukemia; megakaryoblastic leukemia; myeloid sarcoma; and hairy cell leukemia.
In another embodiment for the disclosed method, the cancer is characterized by mutations in the mutS homologues {e.g., MSH2, MSH3, and MSH6), mutL homologues (e.g. MLH1), or mismatch repair endonuclease PMS2. Mutations are changes in the genetic code. They include point mutations and frameshift mutations. In a point mutation, one nucleotide is swapped out for another. Therefore, the mutation occurs at a single point or location within the DNA strand. Frameshift mutations are due to either insertions or deletions of nucleotides. This causes the entire DNA strand to elongate or to shrink in size. Thus, frameshift mutations may alter all of the codons that occur after the deletion or insertion. The mutations referred to herein include, but are not limited to, insertions, deletions, duplications, inversions, or other recognized point mutations. It has now been found that RAD51 inhibitors are particularly effective in treating cancers with mutations in MSH (e.g. MSH6), MLH, or PMS2.
MutS Homo log 2 (MSH2) is a protein that in humans is encoded by the MSH2 gene, which is located on chromosome 2. MSH2 is a tumor suppressor gene and more specifically a caretaker gene that codes for a DNA mismatch repair (MMR) protein, MSH2, which forms a heterodimer with MSH6 to make the human MutSa mismatch repair complex. It also dimerizes with MSH3 to form the MutSP DNA repair complex. MSH2 is involved in many different forms of DNA repair, including transcription-coupled repair, homologous recombination, and base excision repair. Examples of the mutations in MSH2 include, but are not limited to, g.47630253_47630254del, g.47702411_47702421del,
g.47709913_47709915inv, g.47635629_47635634del, g.47637227_47637236dup, g.47639550_47639561del, g.(?_47630206)_(47710367_?)del,
g.(?_47630206)_(47643569_47656880)del, g.47630263_47643568del,
g.(?_47630206)_(4765708 l_47672686)del, g.47630263_47657080del,
g.(?_47630206)_(47672797_47690169)del, g.47630263_47672796del,
g.(?_47630206)_(47672797_47690169)del, g.(?_47630206)_(47693948_47698103)del, g.47630263_47693947del, g.(?_47630206)_(47698202_47702163)del,
g.(?_47630206)_(47630542_47635539)del, g.(?_47630206)_(47708011_47709917)del, g.(?_47630206)_(47635695_47637232)del, g.(?_47630206)_(47635695_47637232)del, g.(?_47630206)_(47637512_47639552)del, g.(?_47630206)_(47639700_47641407)del, g.(?_47630206)_(47641558_47643434)del, g.47618487_47650860delins(155),
g.47628578_47638433del, g.47595033_47662777del, g.47583175_47667707del, g.47625602_47636880del, g.47554933_47699909del, g.47629508_47649552del, g.47629375_47651274del, g.(?_47630206)_(47630542_47635539)del,
g.(?_47630206)_(47635695_47637232)del, g.47643509_476435 lOdel,
g.47643529_47643530dup, g.47656746_47657199dup, g.47656661_47663325del, g.(47643569_47656880)_(47710367_?)del, g.(47643569_47656880)_(47710367_?)del, g.4765688 l_47657080del, g.(47643569_47656880)_(4765708 l_47672686)del, g.(47643569_47656880)_(47657081_47672686)del,
g.(47643569_47656880)_(47657081_47672686)del,
g.(47643569_47656880)_(47657081_47672686)dup,
g.(47643569_47656880)_(47657081_47672686)dup,
g.(47643569_47656880)_(47672797_47690169)del,
g.(47643569_47656880)_(47693948_47698103)del, g.4765688 l_47693947del,
g.(47643569_47656880)_(47702410_47703505)del, g.47656881_47656882ins(173), g.47656901_47656902insA, g.47656903del, g.47656912del, g.47630440del, g.47656923del, g.4765693 l_47656932dup, g.47656943del, g.47656943_47656949delinsCCCAGA, g.47656948dup, g.47656996dup, g.47657000_47657001dup, g.47630449del,
g.47657007dup, g.47657008del, g.47657020_47657023dup, g.47657025_47657026del, g.47657026dup, g.47657030_47657031del, g.47657047_47657050del, g.47657053del, g.47657053_47657057del, g.47657064del, g.47657073dup, g.47657312_47676594del, g.4766861 l_47674615del, g.47672116_47675123del, g.47666463_47677632del, g.47666403_47677572del, g.(47657081_47672686)_(47710367_?)del,
g.(47657081_47672686)_(47710367_?)inv,
g.47671507_47675022delinsCATTCTCTTTGAAAA, g.47657278_47676557del, g.47672687_47672796del, g.(47657081_47672686)_(47672797_47690169)del,
g.(47657081_47672686)_(47672797_47690169)del,
g.(47657081_47672686)_(47693948_47698103)del,
g.(47657081_47672686)_(47698202_47702163)del,
g.(47657081_47672686)_(47708011_47709917)del, g.47672691dup, g.47672697dup, g.47672721_47672744delins47672748_47672771inv, g.47672728_47672729del, g.4767273 ldup, g.47672750_4767275 linsGG, g.47672755_47672758del,
g.47672762_47672763del, g.47630466_47630494del, g.47686194_47697740del, g.(47672797_47690169)_(47710367_?)del,
g.(47672797_47690169)_(47690294_47693796)del,
g.(47672797_47690169)_(47693948_47698103)del, g.47690170_47693947del,
g.(47672797_47690169)_(47693948_47698103)del,
g.(47672797_47690169)_(47693948_47698103)dup,
g.(47672797_47690169)_(47705659_47707834)del, g.47690173del, g.47690191del, g.47690216_47690217dup, g.47690227del, g.47690227dup, g.47690228_47690232del, g.47690230_47690231del, g.47690240del, g.47690240_47690243del, g.47630475del, g.47630475_47630476del, g.47690259_47690260delinsCT, g.47690277dup, g.47690280del, g.47690283dup, g.(47690294_47693796)_(47702410_47703505)del,
g.47630484_47630485insG, g.47693838_47693839del, g.47693862del, g.47693864del, g.47693873del, g.47693880dup, g.47693913del, g.47693924_47693925dup, g.47630493del, g.47697730_47706125del, g.(47693948_47698103)_(47710367_?)del,
g.(47693948_47698103)_(47698202_47702163)del,
g.(47693948_47698103)_(47705659_47707834)del, g.47698107del, g.47698109del, g.47698109_47698110insA, g.47630496del, g.47698118del, g.47698125del,g.47698129dup, g.47698138_47698139del, g.47698142_47698146del, g.47698144dup,
g.47698147_47698148del, g.47698147_47698148dup, g.47698147_47698148insT, g.47698159del, g.47698162del, g.47698506_47703472del, g.47701803_47708848del, g.(47698202_47702163)_(47710367_?)del,
g.(47698202_47702163)_(47702410_47703505)del,
g.(47698202_47702163)_(47703711_47705410)del,
g.(47698202_47702163)_(47705659_47707834)del, g.47702164del,
g.47702175_47702176insA, g.47702183_47702186del, g.47702185_47702186insCT, g.47702190_47702192del, g.47702191dup, g.47702192_47702193del, g.47702213del, g.47702231del, g.47702242dup, g.47702257del, g.47702262_47702263dup,
g.47630516_47630517dup, g.47630517del, g.47630517dup, g.47702289_47702290inv, g.47702293_47702296del, g.47702301dup, g.47702315del, g.47702315del,
g.47702328_47702329del, g.47630522dup, g.47702339del, g.47702371_47702374dup, g.47702384_47702385del, g.47702386_47702389del, g.47702388del,
g.47702388_47702389del, g.47702390del, g.47702390_47702391del,
g.47702400_47702401del, g.47703506_47703710del, g.47703506_47708010del, g.47703510del, g.47703515del, g.47703521_47703522del, g.47703535_47703536del, g.47703546_47703547del, g.47703548_47703611dup, g.47630534del, g.47703571dup, g.47703574_47703581del, g.47703585dup, g.47630350del, g.47632107_47668733del, g.47703613del, g.(47630542_47635539)_(47643569_47656880)del,
g.(47630542_ _47635539)_ _(47643569_ _47656880)inv,
g.(47630542_ _47635539)_ .(47657081_ _47672686)del, g.47635540_47657080del,
g.(47630542_ _47635539)_ _(47672797_ .47690169)del,
g.(47630542_ _47635539)_ _(47690294_ _47693796)del,
g.(47630542_ _47635539)_ _(47705659_ _47707834)del, g.47635540_47635694del,
g.(47630542_ _47635539)_ _(47635695_ _47637232)del,
g.(47630542_ _47635539)_ _(47635695_ _47637232)del, g.(47630542_47635539)_(47637512_47639552)del, g.47703635dup, g.47703641dup, g.47635542_47635549del, g.47703660_47703663del, g.47703667dup, g.47630351dup, g.47703704del, g.47703826_47707938del,
g.(47703711_47705410)_(47705659_47707834)del, g.47705428_47705431del,
g.47705437_47705438insA, g.47635551_47635552del, g.47705440_47705441del, g.47705461del, g.47705490del, g.47705494del, g.47705495del, g.47635557_47635558del, g.47705505del, g.47705535dup, g.47705547del, g.47705560_47705561dup, g.47705561dup, g.47705562dup, g.47705588del, g.47705608_47705609del, g.47705618dup, g.47705627dup, g.47635571_47635601delins(217), g.(47705659_47707834)_(47710367_?)del,
g.(47705659_47707834)_(47708011_47709917)del, g.47707842_47707843del,
g.47707861del, g.47707861_47707874dup, g.47707878_47707884del,
g.47707878_47707884del, g.47707883del, g.47707895_47707905del, g.47707897del, g.47707901_47707902del, g.47707905_47707906del, g.47707921del, g.47635583dup, g.47635583_47635584del, g.47707969_47707973del, g.47707996_47707997ins(l 15), g.47708009_47708010del, g.(47708011_47709917)_(47710367_?)del,
g.47635591_47635592del, g.47635597_47635618dup, g.47635606_47635607del, g.47630359dup, g.47635672del, g.47635675_47635678del, g.47630364dup, g.47635680dup, g.47636862_47639040del, g.47636781_47638831del, g.47636753_47638155del,
g.47636552_47638597del, g.(47635695_47637232)_(47643569_47656880)del,
g.(47635695_ _47637232)_ _(47643569_ _47656880)del,
g.(47635695_ _47637232)_ .(47657081_ _47672686)del,
g.(47635695_ _47637232)_ _(47672797_ .47690169)del,
g.(47635695_ _47637232)_ _(47698202_ _47702163)del,
g.(47635695_ _47637232)_ .(47637512_ _47639552)del,
g.(47635695_ _47637232)_ _(47641558_ _47643434)del, g.47637234del,
g.47637246_47637247del, g.47637253_47637254del, g.47637254_47637255del,
g.47637254_47637255del, g.47637265del, g.47637274del, g.47637282del, g.47637320del, g.47637372_47637375del, g.47637377_47637449dup, g.47637379del, g.47637384del, g.47637394_47637395del, g.47637396_47637397del, g.47637417del,
g.47637427_47637435del, g.47637437_47637439del, g.47637453del, g.47637458dup, g.47637479_47637482dup, g.47637482dup, g.47637504_47637505del,
g.47637508_47637511del, g.47638050_47653430del, g.47638302_47648462del,
g.47638478_47648643del, g.(47637512_47639552)_(47710367_?)del,
g.(47637512_47639552)_(47643569_47656880)del, g.47639553_47643568del, g.(47637512_47639552)_(4765708 l_47672686)del,
g.(47637512_47639552)_(4765708 l_47672686)del,
g.(47637512_47639552)_(47672797_47690169)del,
g.(47637512_47639552)_(47639700_47641407)del,
g.(47637512_47639552)_(47641558_47643434)del, g.47639557_47639561del,
g.47639582_47639586delinsTAAT, g.47639583_47639584del, g.47639594del,
g.47639594dup, g.47639598del, g.47639603_47639604del, g.47639611_47639612del, g.47639612del, g.47639618_47639621del, g.47639624_47639628delinsTTA,
g.47630401dup, g.47639632dup, g.47639638_47639641dup, g.47639638_47639641dup, g.47639639del, g.47639639del, g.47639642dup, g.47630403_47630404insC, g.47639653del, g.47639666del, g.47639666_47639669del, g.47639668del, g.47639670_47639673delinsTT, g.47639674_47639675dup, g.47639695_47639696del, g.47639707_47642985del, g.47641402_47642007del, g.(47639700_47641407)_(47643569_47656880)del,
g.47641408_47643568del, g.(47639700_47641407)_(47657081_47672686)del,
g.(47639700_47641407)_(47672797_47690169)del,
g.(47639700_47641407)_(47641558_47643434)del,
g.(47639700_47641407)_(47641558_47643434)del, g.47641410del,
g.47641425_47641426del, g.47641426_47641429del, g.47630412del, g.47641451del, g.47641454dup, g.47641455dup, g.47641469del, g.47641478del, g.47641488_47641491del, g.47641496_47641497del, g.47641503del, g.47641513_47641514dup,
g.47641530_47641537dup, g.47642509_47655432del,
g.(47641558_47643434)_(47643569_47656880)del,
g.(47641558_47643434)_(47693948_47698103)del, g.47630424_47630433del,
g.47643450dup, g.47643462_47643463del, g.47643462_47643463ins(4),
g.47643464_47643465insNC_000022.10:35788169_35788352, g.47643465dup.
MutS Homo log 3 (MSH3) is a human homologue of the bacterial mismatch repair protein MutS that participates in the mismatch repair (MMR) system. MSH3 typically forms the heterodimer MutSP with MSH2 in order to correct long insertion/deletion loops and base- base mispairs in micro satellites during DNA synthesis. Deficient capacity for MMR is found in approximately 15% of colorectal cancers, and somatic mutations in the MSH3 gene can be found in nearly 50% of MMR-deficient colorectal cancers. Examples of the mutations in MSH3 include, but are not limited to, g.79970809del.
MSH6 encodes MutS homologue 6 (MSH6), a member of the Mutator S (MutS) family of proteins that are involved in DNA mismatch repair (MMR). The MSH6 protein forms a heterodimer with MutS homologue 2 (MSH2) in both human and yeast. Human MSH2/6 recognizes single base-base mismatches and short insertion/deletion loops. Upon recognition of a mismatch, MSH2/6 complex binds and exchanges ADP for ATP, resulting in a conformational change to the complex that precedes base pair dissolution, base excision, and repair.
MSH6 mutations include frameshift and/or nonsense mutations and can result in nonfunctional MSH6 and loss of protein expression. Examples include a frameshift mutation at MSH6 amino acid residue 290 and a compounding missense T1189I.
Inactivating MSH6 mutations can be detected in cancers by routine diagnostics methods. These methods include, but are not limited to, obtaining cancer cells and other diagnostic indicators such as peripheral blood mononuclear cells (PBMCs), PBMC subpopulations, circulating blasts (CD34+ cells), circulating tumor cells and circulating exosomescancer cells by biopsy and blood tests and by obtaining lymphatic or other bodily fluids. It is then determined from the cancer cells or other diagnostic indicators whether the cancer exhibits an inactivating MSH6 mutation is by methodology known in the art, for example, direct DNA sequencing and multiplex ligation dependent probe amplification, RNA sequencing (RNA-Seq), microarray, quantitative PCR, or NanoString™ gene expression panels, or MSH6 protein by immunohistochemistry, flow cytometry, immunocytochemistry or Western blot. Methods for identifying inactivating MSH6 mutations are disclosed in Houlleberghs H, Goverde A, Lusseveld J, Dekker M, Bruno MJ, et al. (2017) Suspected Lynch syndrome associated MSH6 variants: A functional assay to determine their pathogenicity. PLOS Genetics 13(5):
el006765. https://doi.org/10.1371/journal.pgen.1006765.
Examples of the mutations in MSH6 include, but are not limited to,
g.48032846_48032849del, g.48032846_48032849del, g.48032846_48032849del, g.48033337_48033342del, g.48033420_48033422del, g.(?_48010221)_(48034092)del, g.(?_48010221)_(48018263_48023032)del, g.47998510_48020183del,
g.48007276_48020272del, g.48026207del, g.48026223del, g.48026223del,
g.48026257_48026261del, g.48026261_48026265del, g.48026312_48026313del, g.48026398del, g.48026543_48026544dup, g.48026693dup, g.48026702del, g.48026712del, g.48026718dup, g.48026736_48026737delinsAG, g.48026736_48026737delinsG, g.48026750_4802675 ldel, g.48026754_48026757del, g.48026756_48026759del, g.48026759_48026760del, g.48026906del, g.48026928_48026931del, g.48026941dup, g.48026991del, g.48027023_48027024del, g.48027079del, g.48027079_48027082dup, g.48027167_48027168del, g.48027172_48027173dup, g.48027178_48027185del, g.48027184_48027185del, g.48027272_48027275del, g.48027470_48027471del, g.48027501_48027502del, g.48027501_48027502delTG, g.48027657dup,
g.48027691_48027694del, g.48027733_48027736dup, g.48027794_48027796delinsC, g.48027841_48027842del, g.48027887del, g.48027890dup, g.48027973_48027980del, g.48028067del, g.48028098del, g.48028106del, g.48028175_48028176del,
g.48028241_48028242del, g.48028241_48028242delTT, g.48028272_48028284dup, g.48028277_48028278del, g.48030558_48030559del, g.48030126_48032394del, g.48030568del, g.48030581_48030584del, g.48030584_48030585dup, g.48030607del, g.48030645_48030646insT, g.48030647del, g.48030647dup, g.48030649dup, g.48030654_48030660del, g.48030659dup, g.48030697_48030698del, g.48030698del, g.48030706del, g.48030710dup, g.48030727_48030728insC, g.48030765_48030829del, c.3438+797_3438+798insTATinsl839_3439-428,
c.3438+797_3438+798insTATinsl839_3439-428, g.48032121_48032122del, g.48032123_48032124del, g.48032124dup, g.48032126_48032129del,
g.48032129_48032130insA, g.48032129_48032132dup,
g.(48032167_48032756)_(48034092_?)del, g.48032809_48032812del, g.48032835dup, g.48032846_48032849del, g.48033374_48033402dup, g.48033395_48033398del, g.48033421_48033433del, g.48033425_48033428dup, g.48033453_48033454insA, g.48033494_48033523del, g.48033495_48033496del, g.48033593dup,
g.48033610_48033613dup, g.48033629_48033635del, g.48033636_48033639dup, g.48033676_48033682del, g.48033707dup, g.48033709_48033716dup,
g.48033721_48033724dup, g.48033727_48033730dup, g.48033728_48033746dup, g.(48033742_48033743)_(48033742_48033743)ins(32), g.48033746dup,
g.48033748_4803375 ldel, g.48033758_48033768del, g.48033773_48033774insATCA, g.48033773_48033776dup, g.48033785_48033789dup, g.48033887_48033910inv, g.(48018263_48023032)_(48032167_48032756)del,
g.(48018263_48023032)_(48023203_48025749)del, g.48023097_48023098del, g.48025773dup, g.48025832del, g.48025860_48025861insT, g.48025884_48025885del, g.48025967dup.
MutL homo log 1, colon cancer, nonpolyposis type 2 (E. coli) is a protein that in humans is encoded by the MLHl gene located on Chromosome 3. It is a gene commonly associated with hereditary nonpolyposis colorectal cancer. Examples of the mutations in MSH6 include, but are not limited to,
g.37089113_37089115del, g.37089175del, g.37090379_37090393del,
g.37038201_37038202del, g.3704253 l_37042542del, g.37053339_37053355del, g.37053354del, g.37053590_37053591insT, g.37034841_37092337del,
g.(?_37034841)_(37092337_?)del, g.(?_37034841)_(37061955_37067127)del,
g.(?_37034841)_(37035155_37038109)del, g.(?_37034841)_(37035155_37038109)del, g.(?_37034841)_(37070424_37081676)del, g.(?_37034841)_(37083823_37089009)del, g.37034841_37083822del, g.(?_37034841)_(37038201_37042445)del,
g.(?_37034841)_(37042545_37045891)del, g.37034841_37042544del,
g.(?_37034841)_(37042545_37045891)del, g.(?_37034841)_(37042545_37045891)del, g.(?_37034841)_(37045966_37048481)del, g.(?_37034841)_(37050397_37053310)del, g.(?_37034841)_(37059091_37061800)del, g.37034658_37038806del,
g.36961079_37138741del, g.37061923del, g.37061927del, g.37061933del, g.37061939del, g.37061942dup, g.37035140_37035141del, g.37070417del, g.37070417_37070418insT, g.37070419dup, g.37070422_37070423insT, g.37080355_37083368del,
g.(37070424_37081676)_(37092337_?)del,
g.(37070424_37081676)_(37081786_37083758)del,
g.(37070424_37081676)_(37083823_37089009)del, g.37038148_37038151del,
g.37038149del, g.37038149dup, g.37081690_37081691del, g.37081691_37081692del, g.37081706_37081708del, g.37081710_3708171 ldel, g.37035053_37035066del, g.37038154del, g.37038154_37038157del, g.37081738_37081739del, g.37081740del, g.37081753dup, g.37081757_3708176 ldup, g.37081782_37081783insAAGT,
g.37081787_37081793delinsATTT, g.(37081786_37083758)_(37083823_37089009)del, g.(37081786_37083758)_(37089175_37090007)del, g.37083759del, g.37083780dup, g.37083781_37083784del, g.37083781_37083784delCTCA, g.37083808_37083809del, g.37083816del, g.37086069_37089606del, g.37084092_37089247del,
g.37084590_37089786del, g.(37083823_37089009)_(37092337_?)del,
g.(37083823_37089009)_(37089175_37090007)del, g.37089010_37089174del,
g.(37083823_37089009)_(37090509_37091976)del, g.37089023del,
g.37089026_37089027del, g.37089027del, g.37089036del, g.37089036dup, g.37038168dup, g.37089042del, g.37089047del, g.37089050_37089053del, g.37089056_37089057del, g.37089061_37089062del, g.37089078_37089096del, g.37089090dup, g.37089099dup, g.37089107_370891 lOdup, g.37089109_370891 lOdel, g.37089130_37089132del, g.37089130_37089132delAAG, g.37089131delinsTTCTT, g.37089133del, g.37089133delG, g.37089144del, g.37089155del, g.37089155_37089161del, g.37089158_37089161del, g.37089162_37089166del, g.37089171del,
g.(37089175_37090007)_(37090101_37090394)del, g.37035056_37035072del,
g.37090013del, g.37090015dup, g.37038183_37038184del, g.37090024_37090037dup, g.37090025_37090053dup, g.37090027dup, g.37038184dup, g.37090031_37090032insT, g.37090041del, g.37090057del, g.37090064_37090067del, g.37038188del, g.37090082del, g.37090086_37090087del, g.37090087_37090088del, g.37090097_37090101delinsC, g.37090099del, g.37038191dup, g.(37090101_37090394)_(37092337_?)del,
g.37035057_37035073del, g.37090405dup, g.37090411_37090415del, g.37090414del, g.37038194del, g.37038198del, g.37090472_37090478del, g.37039445_37059613dup, g.37039760_37052440del, g.3709048 l_37090482del, g.37090483_37090484del, g.37090483_37092045del, g.37040732_37043185delinsACATAGTA,
g.37042445_37042446del, g.(37038201_37042445)_(37042545_37045891)del,
g.(37038201_37042445)_(37048555_37050304)del,
g.(37038201_37042445)_(37050397_37053310)del,
g.(37038201_37042445)_(37053591_37055922)del, g.37090497_37090498del,
g.37090497_37090498delTC, g.37090504_37090507del,
g.(37090509_37091976)_(37092337_?)del, g.(37090509_37091976)_(37092337_?)dup, g.37091977_37091978del, g.37091978_37091987del, g.37042448_37042451del, g.37091984_37091990del, g.37042451_37042453del, g.37092020_37092021del, g.37092022_37092068dup, g.37092027_37092028del, g.37092027_37092028dup, g.37092030dup, g.37092052_37092055del, g.37092054_37092055del,
g.37092068_37092071dup, g.37092091dup, g.37092094_37092097delins(30),
g.37092096_37092106del, g.37092097del, g.37092125_37092126delAA,
g.37092125_37092126del, g.37092139_37092142dup, g.37092142dup, g.37035060dup, g.37042469_37042470del, g.37042470del, g.37042482dup, g.37042485del, g.37042499del, g.37042546dup, g.37044472_37046589del, g.37045648_37049941del,
g.37045095_37054651del, g.37045072_37046861del,
g.(37042545_37045891)_(37045966_37048481)del,
g.(37042545_37045891)_(37092337_?)del,
g.(37042545_37045891)_(37048555_37050304)del,
g.(37042545_37045891)_(37050397_37053310)del, g.37045892_37050396del,
g.37035069del, g.37045926del, g.37045931del, g.37045939_37045940dup,
g.37045957_37045958del, g.37045963del, g.37035075del, g.37048067_37049287del, g.(37045966_37048481)_(37048555_37050304)del,
g.(37045966_37048481)_(37050397_37053310)del, g.37048483del,
g.37048483_37048503delinsT, g.37048486_37048487delinsGTT, g.37048489del, g.37048490del, g.37035076_37035077insCCCA, g.37035077_37035078dup,
g.37048505_37048508del, g.37048521del, g.37048529dup, g.37035082dup,
g.37049873_3705228 ldel, g.37049839_37052249del, g.37049800_37052209del,
g.37049640_37050445del, g.37050305_37050396del,
g.(37048555_37050304)_(37050397_37053310)del, g.37050305_37050396del,
g.37050319_37050320del, g.37050339del, g.37050348del, g.37050353_37050354del, g.37050354dup, g.37050364del, g.37050375_37050376insGA, g.37035090del,
g.37050382_37050383delinsAT, g.37050382_37050383delinsCT, g.37050390_37050396del, g.37052950_37060990del, g.(37050397_37053310)_(37067499_37070274)dup,
g.(37050397_37053310)_(37053591_37055922)del,
g.(37050397_37053310)_(37056036_37058996)del, g.37053353del,
g.37053510_37053511del, g.37035099del, g.37053545_37053546insT, g.37053562del, g.37053578del, g.37053578dup, g.37053585del, g.37053586_37053589del, g.37053591del, g.37053590_37053591delinsAT, g.37055920_37055921del, g.37055914_37055938del, g.(37053591_37055922)_(37070424_37081676)del,
g.(37053591_37055922)_(37083823_37089009)del,
g.(37053591_37055922)_(37059091_37061800)del, g.37035105del, g.37055928dup, g.37035106_37035116del, g.37055938del, g.37035108del, g.37055972_37055975del, g.37055976_37055979del, g.37035111del, g.37055990dup, g.37035114del, g.37035116del, g.37056036del, g.37056037dup, g.37058993_37059001del,
g.(37056036_37058996)_(37070424_37081676)del,
g.(37056036_37058996)_(37059091_37061800)del, g.37058997_37059000del,
g.37059014_37059017del, g.37059017_37059021del, g.37059027_37059030dup, g.37035122del, g.37059062_37059063insT, g.37059065_37059066del, g.37059066del, g.37059066dup, g.37059072_37059073del, g.37059072_37059073dup,
g.37059090_37059093del, g.37061595_37061913del, g.37061308_37066756del,
g.37061207_37063077del, g.(37059091_37061800)_(37092337_?)del,
g.(37059091_37061800)_(37061955_37067127)del, g.37061801_37061954del,
g.(37059091_37061800)_(37083823_37089009)del, g.37061803dup, g.37061804del, g.37061817del, g.37061837_37061838dup, g.37061844del, g.37061851dup, g.37061855dup, g.37061870del, g.37061904_37061906del, g.37061910del, g.37035047del,
g.[37049179_37051317delinsTG;37051667_37054327delinsCA] .
Human PMS2 related genes are located at bands 7pl2, 7pl3, 7ql 1, and 7q22. Exons 1 through 5 of these homologues share high degree of identity to human PMS2. The product of this gene is involved in DNA mismatch repair. The protein forms a heterodimer with MLHl and this complex interacts with MSH2 bound to mismatched bases. Defects in this gene are associated with hereditary nonpolyposis colorectal cancer, with Turcot syndrome, and are a cause of supratentorial primitive neuroectodermal tumors.
Examples of the mutations in PMS2 include, but are not limited to,
g.(?_6012870)_(6048737_?)del, g.6012870_6048737del,
g.(6027252_6029430)_(6048737_?)del, g.(6045663_6048627)_(6048737_?)del,
g.6029554del, g.6029499dup, g.6029495_6029496del, g.6029462_6029463delinsTAAA, g.5992485_6028601del, g.(6018328_6022454)_(6027252_6029430)del,
g.(6013174_6017218)_(6027252_6029430)del, g.6027226_6027227ins(20), g.6027175del, g.6027090dup, g.6036705_6044207delinsCG, g.6026666dup, g.6026628del, g.6043671del, g.6026565dup, g.6026565dupT, g.6018315_6018316del, g.6018306_6018310del, g.6018306_6018310delAGTTA, g.6043633_6043634dup, g.6018256_6018259del, g.6015623_6017501del, g.6016429_6017479del, g.6017300_6017303del,
g.6045579_6045674delinsATTT, g.(6043690_6045522)_(6045663_6048627)del, g.(?_6012870)_(6042268_6043320)del, g.(6035265_6036956)_(6042268_6043320)del, g.6038283_6039384del, g.6038901del, g.6038851dup,
g.(6035265_6036956)_(6037055_6038738)del,
g.6037019_6037024delinsCTTCACACACA, g.6036980del, g.6036958dup,
g.6035323_6035324insJN866832.1, g.(6022623_6026389)_(6035265_6036956)del, g.(6031689_6035164)_(6035265_6036956)del, g.6035204_6035207del,
g.6035205_6035206del, g.(?_6012870)_(6031689_6035164)del,
g.(6027252_6029430)_(6031689_6035164)del,
g.(6029587_6031603)_(6031689_6035164)del, g.6028725_6029882del,
g.(?_6012870)_(6029587_6031603)del.
The present invention provides a method of treating patients with Lynch syndrome to reduce the likelihood of from developing or treating cancers derieved from Lynch syndrome, by administering to the subject an effective amount of one or more disclosed compounds, or a pharmaceutically acceptable salt thereof, or the corresponding pharmaceutical composition. Lynch syndrome is a hereditary disorder caused by a mutation in a mismatch repair gene in which affected individuals have a higher than normal chance of developing colorectal cancer, endometrial cancer, and various other types of aggressive cancers, often at a young age - also called hereditary nonpolyposis colon cancer (HNPCC).
The mutations of specific mismatch repair (MMR) genes including but not limited to MLH1, MSH2, MSH6, PMS2, and EPCAM-TACSTD 1 deletions are responsible for Lynch syndrome. These genes work in repairing mistakes made when DNA is copied in preparation for cell division. The defects in the genes disallow repair of DNA mistakes and as cells divide, errors stack and uncontrollable cell growth may result in cancer.
Those with Lynch syndrome carry up to an 85% risk of contracting colon cancer as well as a higher than average risk for endometrial cancer, stomach cancer, pancreatic cancer, kidney/ureter tract cancer, hepatobiliary tract cancer, gastric tract cancer, prostate cancer, ovarian cancer, gallbladder duct cancer, brain cancer, small intestine cancer, breast cancer, and skin cancer.
Thus, in one embodiment for the disclosed method, the method is a method of treating cancer derived from Lynch syndrome, selected from the group consisting of colon cancer, endometrial cancer, stomach cancer, pancreatic cancer, kidney/ureter tract cancer, hepatobiliary tract cancer, gastric tract cancer, prostate cancer, ovarian cancer, gallbladder duct cancer, brain cancer, small intestine cancer, breast cancer, and skin cancer.
In yet another embodiment, the method is a method of treating autoimmune disease. Exemplary autoimmune diseases include lupus erythematosus; Wiskott-Aldrich syndrome; autoimmune lymphoproliferative syndrome; myasthenia gravis; rheumatoid arthritis (RA); lupus nephritis; multiple sclerosis; systemic lupus erythematosis; discoid lupus; subacute cutaneous lupus erythematosus; cutaneous lupus erythematosus including chilblain lupus erythematosus; chronic arthritis; Sjogren's syndrome; inflammatory chronic rhino sinusitis; colitis; celiac disease; inflammatory bowel disease; Barrett's esophagus; inflammatory gastritis; autoimmune nephritis; autoimmune vasculitis; autoimmune hepatitis; autoimmune carditis; autoimmune encephalitis; autoimmune diabetes; autoimmune diabetes nephritis; psoriasis; Graft-versus-host disease (GvHD); and autoimmune mediated hematological disease.
In one aspect of this embodiment, the method is a method of treating immune deficiency selected from the group consisting of Autoimmune Lymphoproliferative
Syndrome (ALPS), Autoimmune polyglandular syndrome type 1 (APS-1), BENTA Disease, Caspase Eight Deficiency State (CEDS), Chronic Granulomatous Disease (CGD), Common Variable Immunodeficiency (CVID), Congenital Neutropenia Syndromes, CTLA4
Deficiency, DOCK8 Deficiency, GATA2 Deficiency, Glycosylation Disorders With
Immunodeficiency, hyper-immunoglobulin E syndrome (HIES), Hyper-Immunoglobulin M (Hyper-IgM) Syndromes, Leukocyte adhesion deficiency (LAD), LRBA deficiency, PI3 Kinase disease, PLCG2-associated antibody deficiency and immune dysregulation (PLAID), severe combined immunodeficiency (SCID), STAT3 gain-of-function disease, Warts, Hypogammaglobulinemia, Infections, and Myelokathexis Syndrome (WHIMS), X-Linked Agammaglobulinemia (XLA), X-Linked Lymphoproliferative Disease (XLP), and XMEN Disease.
As used herein, the term "immune deficiency" refers to a condition in which a portion or some portions of cell components constituting an immune system are defective or dysfunction, so that a normal immune mechanism is damaged. In other words, "immune deficiency" means a condition under which: congenital immunity and/or acquired immunity are suppressed and/or decreased. In some embodiments, the immune -deficiency subject is an immunocompromised subject. Non-limiting examples of immune deficiencies can include AIDS, hypogammaglobulinemia, agammaglobulinemia, granulocyte deficiency, chronic granulomatous disease, asplenia, SCID, complement deficiency, and/or sickle cell anemia.
In another aspect of this embodiment, the method is a method of treating a neurodegenerative disorder selected from the group consisting of multiple sclerosis, Parkinson's disease (PD), Alzheimer's disease (AD), Dentatorubropallidoluysian atrophy (DRPLA), Huntington's Disease (HD), Spinocerebellar ataxia Type 1 (SCA1),
Spinocerebellar ataxia Type 2 (SCA2), Spinocerebellar ataxia Type 3 (SCA3),
Spinocerebellar ataxia 6 (SCA6), Spinocerebellar ataxia Type 7 (SCA7), Spinocerebellar ataxia Type 8 (SCA8), Spinocerebellar ataxia Type 12 (SCA12), Spinocerebellar ataxia Type 17 (SCA17), Spinobulbar Muscular Ataxia/Kennedy Disease (SBMA), Fargile X syndrome (FRAXA), Fragile XE mental retardation (FRAXE), and Myotonic dystrophy (DM).
A "subject" is a mammal, preferably a human, but can also be an animal in need of veterinary treatment, e.g. , companion animals (e.g. , dogs, cats, and the like), farm animals (e.g., cows, sheep, pigs, horses, and the like) and laboratory animals (e.g. , rats, mice, guinea pigs, and the like).
In certain embodiments, the methods disclosed herein further comprise coadministering an effective amount of a DNA repair inhibitor, a DNA damage response (DDR) inhibitor, a DNA damaging agent or an immunomodulatory agent to the subject being treated for cancer, in addition to an effective amount of a disclosed RAD51 inhibitor. The term "DNA repair inhibitor" refers to any agent that targets
components/processes which a cell uses to repair mutations or changes in DNA and restore the DNA to its original state and prevents the repair of DNA. Examples of DNA repair inhibitors include: RPA inhibitors, APE1 inhibitors, DNA ligase inhibitors, DNA polymerase inhibitors, Parp inhibitors etc.
The term "DNA damage response inhibitor" refers to any agent that targets components/processes involved in detecting DNA lesions, signaling the presence of DNA damage, and/or promote the repair of DNA damage. Examples of DNA damage response inhibitors include checkpoint inhibitors, ATM and ATR inhibitiors, DNA-PK inhibitors, etc.
The term "DNA damaging agent" refers to any agent that directly or indirectly damages DNA for which homologous recombination could repair the damage. The DNA damaging agents is selected from the group consisting of: exposure to a DNA damaging chemical; exposure to a chemotherapeutic agent; exposure to a radiochemotherapy, and exposure to ionizing or ultraviolet radiation. Specific examples of DNA-damaging chemotherapeutic agents include alkylating agents, nitrosoureas, anti-metabolites, plant alkaloids, plant extracts and radioisotopes. Specific examples of the chemotherapeutic agents also include DNA-damaging drugs, for example, 5-fluorouracil (5-FU), capecitabine, S- l (Tegafur, 5-chloro-2,4-dihydroxypyridine and oxonic acid), 5-ethynyluracil, arabinosyl cytosine (ara-C), 5-azacytidine (5-AC), 2',2'-difluoro-2'-deoxycytidine (dFdC), purine antimetabolites (mercaptopurine, azathiopurine, thioguanine), gemcitabine hydrochlorine (Gemzar), pentostatin, allopurinol, 2- fluoro- arabinosyl- adenine (2F-ara-A), hydroxyurea, sulfur mustard (bischloroetyhylsulfide), mechlorethamine, melphalan, chlorambucil, cyclophosphamide, ifosfamide, thiotepa, AZQ, mitomycin C, dianhydrogalactitol, dibromoducitol, alkyl sulfonate (busulfan), nitrosoureas (BCNU, CCNU, 4-methyl CCNU or ACNU), procarbazine, decarbazine, rebeccamycin, anthracyclins such as doxorubicin (adriamycin; ADR), daunorubicin (Cerubicine), idarubicin (Idamycin) and epirubicin (Ellence), anthracyclin analogs such as mitoxantrone, actinimycin D, non-intercalating topoisomerase inhibitors such as epipodophyllotoxins (etoposide or VP16, teniposide or VM- 26), podophylotoxin, bleomycin (Bleo), pepleomycin, compounds that form adducts with nucleic acid including platinum derivatives, e.g. , cisp latin (CDDP), trans analog of cisp latin, carboplatin, iproplatin, tetraplatin and oxaliplatin, as well as camptothecin, topotecan, irinotecan (CPT- 11), and SN-38. Specific examples of nucleic acid damaging treatments include radiation e.g. , ultraviolet (UV), infrared (IR), or .alpha.-, .beta.-, or .gamma.- radiation, as well as environmental shock, e.g., hyperthermia. "Immunomodulatory agent" means an agent that modulates an immune response to an antigen but is not the antigen or derived from the antigen. "Modulate", as used herein, refers to inducing, enhancing, suppressing, directing, or redirecting an immune response. Such agents include immuno stimulatory agents, such as adjuvants, that stimulate (or boost) an immune response to an antigen but is not an antigen or derived from an antigen. There are several distinct types of immunomodulatory agents, which include, but are not limited to, Toll-like Receptor (TLR) agonists and Toll-like Receptor (TLR) antagonists. Such agents also include immunosuppressants. The immunomodulatory agent is selected from the group consisting of immune checkpoint modulators, Toll-like receptor (TLR) agonists, cell-based therapies, cytokines and cancer vaccines.
In certain embodiments, the subject is determined to have an increased level and/or activity of a DNA damage process or DNA editing enzyme. In one aspect of this
embodiment, the DNA editing enzyme is selected from the group consisting of activation induced cytidine deaminase (AID or AICDA), APOBEC2, APOBEC3A, APOBEC3C, APOBEC3D, APOBEC3F, APOBEC3G, APOBEC3H, APOBEC4, a Type 1 Topoisomerase, a Type 2 Topoisomerase, Recombination Activating Gene 1 (RAG 1), and Recombination Activating Gene 2 (RAG2).
In certain embodiments, blood cells obtained from the subject have been determined to have a detectable level of activation- induced cytidine deaminase (AID).
In certain embodiments, B cells obtained from the subject have been determined to have a detectable level of activation- induced cytidine deaminase (AID).
In certain embodiments, the detectable level of activation- induced cytidine deaminase (AID) is statistically significantly higher than the level of AID expressed in unactivated B- cells or normal non- immune cells from a healthy subject.
Methods of Administration and Dosage Forms
The precise amount of compound administered to provide an "effective amount" to the subject will depend on the mode of administration, the type, and severity of the disease, and on the characteristics of the subject, such as general health, age, sex, body weight, and tolerance to drugs. The skilled artisan will be able to determine appropriate dosages depending on these and other factors. When administered in combination with other therapeutic agents, e.g., when administered in combination with an anti-cancer agent, an "effective amount" of any additional therapeutic agent(s) will depend on the type of drug used. Suitable dosages are known for approved therapeutic agents and can be adjusted by the skilled artisan according to the condition of the subject, the type of condition(s) being treated and the amount of a compound of the invention being used by following, for example, dosages reported in the literature and recommended in the Physician 's Desk Reference (57th ed., 2003).
The term "effective amount" means an amount when administered to the subject which results in beneficial or desired results, including clinical results, e.g., inhibits, suppresses or reduces the symptoms of the condition being treated in the subject as compared to a control. For example, a therapeutically effective amount can be given in unit dosage form (e.g., 0.1 mg to about 50 g per day, alternatively from 1 mg to about 5 grams per day).
The terms "administer", "administering", "administration", and the like, as used herein, refer to methods that may be used to enable delivery of compositions to the desired site of biological action. These methods include, but are not limited to, intraarticular (in the joints), intravenous, intramuscular, intratumoral, intradermal, intraperitoneal, subcutaneous, orally, topically, intrathecally, inhalationally, transdermally, rectally, and the like.
Administration techniques that can be employed with the agents and methods described herein are found in e.g., Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.
In addition, the disclosed RAD51 inhibitors can be co-administered with other therapeutic agents. As used herein, the terms "co-administration", "administered in combination with", and their grammatical equivalents, are meant to encompass
administration of two or more therapeutic agents to a single subject, and are intended to include treatment regimens in which the agents are administered by the same or different route of administration or at the same or different times. In some embodiments the one or more compounds described herein will be co- administered with other agents. These terms encompass administration of two or more agents to the subject so that both agents and/or their metabolites are present in the subject at the same time. They include simultaneous administration in separate compositions, administration at different times in separate compositions, and/or administration in a composition in which both agents are present. Thus, in some embodiments, the compounds described herein and the other agent(s) are
administered in a single composition. In some embodiments, the compounds described herein and the other agent(s) are admixed in the composition.
The particular mode of administration and the dosage regimen will be selected by the attending clinician, taking into account the particulars of the case (e.g., the subject, the disease, the disease state involved, the particular treatment). Treatment can involve daily or multi-daily or less than daily (such as weekly or monthly etc.) doses over a period of a few days to months, or even years. However, a person of ordinary skill in the art would immediately recognize appropriate and/or equivalent doses looking at dosages of approved compositions for treating a a RAD51 mediated disease using the disclosed RAD51 inhibitors for guidance.
The compounds or the corresponding pharmaceutical compositions taught herein can be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. The compounds of the present teachings may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump or transdermal administration and the pharmaceutical compositions formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal and topical modes of administration. Parenteral administration can be by continuous infusion over a selected period of time.
The pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration. In an embodiment, the composition is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous, subcutaneous, intramuscular, oral, intranasal, or topical administration to human beings. In preferred embodiments, the pharmaceutical composition is formulated for intravenous administration.
Typically, for oral therapeutic administration, a compound of the present teachings may be incorporated with excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
Typically for parenteral administration, solutions of a compound of the present teachings can generally be prepared in water suitably mixed with a surfactant such as hydro xypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
Typically, for injectable use, sterile aqueous solutions or dispersion of, and sterile powders of, a compound described herein for the extemporaneous preparation of sterile injectable solutions or dispersions are appropriate. EXEMPLIFICATION
Abbreviations:
Ac acetyl
ACN acetonitrile
aq aqueous
Bn benzyl
Boc ie/t-butoxycarbonyl
br. broad
d doublet (only when used within 1H NMR spectra)
DCM dichloromethane
DIEA(DIPEA) diisopropylethylamine
DMA dimethylacetamide
DMAP 4-dimethylaminopyridine
DMF N,N-dimethylformamide
DMSO dimethylsulfoxide
dppf 1, 1'- bis( diphenylphosphino) ferrocene
eq equivalent
EtOAc ethyl acetate
hr hour
HBTU N,N,N',N',-tetramethyl-0-(lH-benzotriazol- l-yl)uronium hexafluoropho sphate
HPLC high performance liquid chromatography
LC-MS liquid chromatography coupled to mass spectrometry m multiplet
MS ESI mass spectra, electrospray ionization
NBS N-bromosuccinimide
NMR nuclear magnetic resonance
prep preparative
Py pyridine
s singlet
sat saturated
SFC supercritical fluid chromatography
t triplet TEA triethylamine
TFA trifluoro acetic acid
THF tetrahydrofuran
TLC thin layer chromatography
Tol toluene
Figure imgf000055_0002
Example 1. Synthesis of (S)-(l-methylpyrrolidin-2-yl)methyl (4-(2-(4- ((benzylcarbamoyl) oxy)piperidin-l-yl)thiazol-5-yl)-3-(N-(tert- butyl)sulfamoyl)phenyl)carbamate
Scheme 1:
Figure imgf000055_0001
HCI/MeOH
Figure imgf000056_0001
14
Figure imgf000056_0002
15 16
Figure imgf000056_0003
Ex. 1
General method D for preparation of sulfonamide compound 12.
Figure imgf000056_0004
11 12
To a solution of 5-amino-2-(2-bromothiazol-5-yl)-N-tert-butyl-benzenesulfonamide (5 g, 12.8 mmol, 1 eq.) in DCM (30 mL) were added DMAP (156 mg, 1.3 mmol, 0.1 eq.) and Ac20 (1.96 g, 19.2 mmol, 1.5 eq.). The mixture was stirred at 20°C for 1 hr, and then washed with 1M HC1 (50 mL) and sat.aq.Na2C03 (50 mL). The organic layer was dried over Na2S04, filtered and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether: Ethyl acetate = 5: 1 to 2: 1) to give N-[4-(2-bromothiazol-5-yl)-3-(tert- butylsulfamoyl)phenyl]acetamide (1.5 g, 3.5 mmol, 27% yield) as a yellow solid. ESI [M+H] = 433.9/431.9
General method A for preparation of sulfonamide compound 13.
Figure imgf000056_0005
12 13 To a solution of N-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl)phenyl]acetamide (700 mg, 1.6 mmol, 1 eq.) in DMF (20 mL) were added K2CO3 (448 mg, 3.3 mmol, 2 eq.) and piperidin-4-ol (246 mg, 2.4 mmol, 1.5 eq.). The mixture was stirred at 100°C for 12 hrs and then poured into H20 (100 mL). The aqueous phase was extracted with EtOAc (50 mL x 3), the combined organic layers were washed with brine (100 mL), dried over Na2S04, filtered and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether: Ethyl acetate = 1 :2) to give N-[3-(tert-butylsulfamoyl)-4-[2-(4-hydroxy-l- piperidyl)thiazol-5-yl]phenyl]acetamide (550 mg, 1.2 mmol, 13.2% yield) as a yellow solid. 1H NMR (400MHz, CHLOROFORM-d) δ = 8.26 (dd, J=2.2, 8.6 Hz, 2H), 7.93 (d, J=2.4 Hz, 1H), 7.37 (d, J=8.4 Hz, 1H), 7.31 (s, 1H), 4.02 - 3.94 (m, 1H), 3.88 - 3.80 (m, 2H), 3.30 (ddd, J=3.5, 9.2, 13.1 Hz, 2H), 2.21 (s, 3H), 2.03 - 1.95 (m, 2H), 1.67 (dtd, J=4.0, 8.7, 13.0 Hz, 2H), 1.09 (s, 9H). ESI [M+H] = 453.1
General method F for preparation of sulfonamide compound 15.
Figure imgf000057_0001
14 15
[l-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl] N- benzylcarbamate (380 mg, 649 umol, 1 e .) was dissolved into HCl/MeOH (4 M, 20 mL) and the mixture was stirred at 20°C for 1 hr. The mixture was concentrated, diluted with sat.aq.Na2C03 (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na2S04, filtered and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether: Ethyl acetate = 10: 1 to 1 : 1) to give [l-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl] N- benzylcarbamate (300 mg, 552 umol, 85% yield) as a yellow solid. ESI [M+H] = 544.2
General method Hfor preparation of Example 1.
Figure imgf000057_0002
16 Ex. 1 To a solution of [(2S)- l-methylpyrrolidin-2-yl]methanol (42 mg, 367 umol, 2 eq.) and DIEA (71.11 mg, 550 umol, 3 eq.) in MeCN (5 mL) was added a solution of [l-[5-[2-(tert- butylsulfamoyl)-4-[ ( 4-nitrophenoxy )carbonylamino ]phenyl]thiazol-2-yl]-4-piperidyl] N- benzylcarbamate (130 mg, 184 umol, 1 eq.) in DCM (2 mL) and the mixture was refluxed for 1 hr. The mixture was concentrated and the residue was purified by prep-HPLC to give (S)- ( l-methylpyrrolidin-2-yl)methyl ( 4-(2-( 4-( (benzylcarbamoyl)oxy )piperidin-l-yl)thiazol-5- yl)-3-(N-(tert-butyl)sulfamoyl)phenyl)carbamate (19.87 mg, 28.8 umol, 15.7% yield, 99.3% purity) as a pale yellow solid. 1H NMR (400MHz, DMSO-d6) δ = 10.06 (s, 1H), 8.26 (d, J=2.0 Hz, 1H), 7.71 (t, J=6.2 Hz, 1H), 7.60 (dd, J=2.3, 8.5 Hz, 1H), 7.37 - 7.14 (m, 7H), 6.94 (s, 1H), 4.81 - 4.71 (m, 1H), 4.16 (br d, J=6.2 Hz, 2H), 4.10 - 4.03 (m, 1H), 4.02 - 3.96 (m, 1H), 3.73 - 3.63 (m, 2H), 3.28 (br s, 2H), 2.94 - 2.87 (m, 1H), 2.44 - 2.37 (m, 1H), 2.30 (s, 3H), 2.18 - 2.05 (m, 1H), 1.99 - 1.82 (m, 3H), 1.68 - 1.52 (m, 5H), 1.12 - 1.03 (m, 9H). ESI [M+H] = 685.2
Example 2. Synthesis of [l-[5-[2-(tert-butylsulfamoyl)-4-[[(2R)-l-methylpyrrolidin-2- yl]methoxycarbonylamino]phenyl]thiazol-2-yl]-4-piperidyl] N -benzylcarbamate.
The following compound was synthesized via same method by the key intermediate 16.
Figure imgf000058_0001
Ex. 2
[l-[5-[2-(tert-butylsulfamoyl)-4-[[(2R)-l-methylpyrrolidin-2- yl]methoxycarbonylamino]phenyl]thiazol-2-yl]-4-piperidyl] N -benzylcarbamate . 1H NMR
(400MHz, DMSO-d6) δ = 10.07 (s, 1H), 8.29 (d, J=2.2 Hz, 1H), 7.73 (t, J=6.2 Hz, 1H), 7.63 (dd, J=2.3, 8.5 Hz, 1H), 7.39 - 7.19 (m, 7H), 6.94 (s, 1H), 4.80 (td, J=4.1, 8.0 Hz, 1H), 4.20 (br d, J=6.1 Hz, 2H), 4.14 - 4.07 (m, 1H), 4.06 - 3.99 (m, 1H), 3.72 (br d, J=13.6 Hz, 2H), 3.29 - 3.22 (m, 2H), 2.98 - 2.92 (m, 1H), 2.45 - 2.41 (m, 1H), 2.33 (s, 3H), 2.22 - 2.13 (m, 1H), 2.02 - 1.79 (m, 3H), 1.72 - 1.56 (m, 5H), 1.20 - 1.03 (m, 9H). ESI [M+H] = 685.2 Example 3. Synthesis of [l-[5-[4-(benzylcarbamoylamino)-2-(tert- butylsulfamoyl)phenyl] thiazol-2-yl]-4-piperidyl] N-isopropylcarbamate.
Scheme 2:
Figure imgf000059_0001
22 23
Figure imgf000059_0002
Ex. 3
Preparation of compound 23.
Figure imgf000059_0003
22 23
General method A, l-benzyl-3-[3-(tert-butylsulfamoyl)-4-[2-(4-hydroxy-l-piperidyl) thiazol- 5-yl]phenyl]urea. ESI [M+H] = 544.1
Preparation of Ex. 3.
Figure imgf000059_0004
23 Ex. 3
To a solution of l-benzyl-3-[3-(tert-butylsulfamoyl)-4-[2-(4-hydroxy-l-piperidyl) thiazol-5- yl]phenyl]urea (50 mg, 91.96 umol, 1 eq.) in Tol. (2 mL), were added DMAP (22.47 mg, 183.92 umol, 2 eq.) and [isopropyl(methyl)-azanylidene] methanone (78.26 mg, 919.62 umol, 10 eq.). The mixture was stirred at 100°C for 4 hrs and then concentrated. The residue was purified by prep-TLC (Petroleum ether/EtOAc=l :3) and then acidic prep-HPLC to give [1- [5-[4-(benzylcarbamoylamino )-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl] N- isopropylcarbamate (28.31 mg , 99.2% purity) as a yellow solid. 1H NMR (400MHz, METHANOL-d4) δ = 8.29 (d, J=2.0 Hz, 1H), 7.71 (dd, J=2.0, 8.3 Hz, 1H), 7.43 (d, J=8.8 Hz, 1H), 7.35 (d, J=3.9 Hz, 5H), 7.27 (dt, J=2.7, 5.7 Hz, 1H), 4.98 - 4.93 (m, 1H), 4.43 (s, 2H), 3.90 - 3.59 (m, 5H), 2.19 - 2.07 (m, 2H), 1.99 - 1.86 (m, 2H), 1.22 (s, 9H), 1.16 (d, J=6.8 Hz, 6H). ESI [M+H] = 629.1
Example 4. Synthesis of isopropyl 4-[5-[2-(tert- butylsulfamoyl)-4-(2-pyridylmethyl carbamoylamino)phenyl]thiazol-2-yl]piperidine-l-carboxylate.
Scheme 3:
Figure imgf000060_0001
Ex. 4
Preparation of compound 45
Figure imgf000060_0002
General method D, 4-nitrophenyl ( 4-bromo-3-(N-( tert-butyl)sulfamoyl)phenyl) carbamate. ESI [M+H] =474.1
General method Efor preparation of sulfonamide compound 46.
Figure imgf000061_0001
To a solution of 2-pyridylmethanamine (352 mg, 3.3 mmol, 5 eq.) and DIEA (84 mg, 650 umol, 1 eq.) in DCM (3 mL) was added the solution of (4-nitrophenyl) N-[4-bromo-3-(tert- butylsulfamoyl)pheny I] carbamate (307 mg, 650 umol, 1 eq.) in DCM (3 mL). The mixture was stirred at 25°C for 1 hr, then diluted with DCM (30 mL) and washed with H20 (20 mL x 2). The organic layer was concentrated and the residue was purified by prep-TLC (Si02, Petroleum ether: Ethyl acetate = 0: 1) to give l-[4-bromo-3-(tert-butylsulfamoyl) phenyl]-3- (2-pyridylmethyl)urea (180 mg, crude) as a yellow solid. ESI [M+H] =441.2/443.2
Preparation of compound 47
Figure imgf000061_0002
A mixture of l-[4-bromo-3-(tert-butylsulfamoyl)phenyl]-3-(2-pyridylmethyl)urea (140 mg, 317.21 umol, 1 eq.), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl- l,3,2- dioxaborolan-2-yl)- 1,3,2-dioxaborolane (322.21 mg, 1.27 mmol, 4 eq.), Pd(dppf)Cl2 (116.05 mg, 158.61 umol, 0.5 eq.) and KOAc (93.39 mg, 951.64 umol, 3 eq.) in dioxane (4 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 80°C for 12 hrs under N2 atmosphere. The mixture was concentrated and the residue was purified by column chromatography (Si02, Petroleum ether/Ethyl acetate=l/0 to 1 : 1) to give l-[3-(tert- butylsulfamoyl) -4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)phenyl]-3-(2- pyridylmethyl)urea (50 mg, 89.37 umol, 28.17% yield, 87.3% purity). ESI [M+H] =489.4 General method K for preparation of compound Ex. 4
Figure imgf000062_0001
47 Ex. 4
A mixture of l-[3-(tert-butylsulfamoyl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan -2- yl)phenyl]-3-(2-pyridylmethyl)urea (26.38 mg, 54.01 umol, 1.2 eq. ), isopropyl 4-(5- bromothiazol-2-yl)piperidine-l-carboxylate (15 mg, 45.01 umol, 1 eq. ), Na2C03 (9.54 mg, 90.02 umol, 2 eq. ), KF (7.85 mg, 135.04 umol, 3 eq. ) and Pd(PPh3)4 (5.20 mg, 4.50 umol, 0.1 eq. ) in H20 (0.1 mL)/EtOH (0.3 mL)/Tol. (0.3 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 85°C for 12 hrs under N2 atmosphere. The reaction mixture was filtered and concentrated. The residue was purified by prep-TLC (Si02, Petroleum ether/Ethyl acetate=l/2) and then prep-HPLC (TFA condition) to give isopropyl 4- [5-[2-(tert- butylsulfamoyl)-4-(2-pyridylmethylcarbamoylamino)phenyl]thiazol-2- yl]piperidine-l-carboxylate (6.36 mg, 8.21 umol, 18.24% yield, 94.07% purity, TFA) as a pale yellow solid. 1H NMR (400MHz, METHANOL-d4) δ = 8.75 (br d, J=5.6 Hz, IH), 8.53 (br t, J=7.9 Hz, IH), 8.34 (d, J=2.0 Hz, IH), 8.04 (br d, J=8.1 Hz, IH), 7.92 (br t, J=6.5 Hz, IH), 7.75 (s, IH), 7.69 (dd, J=2.3, 8.4 Hz, IH), 7.39 (d, J=8.3 Hz, IH), 4.94 - 4.92 (m, IH), 4.77 (s, 2H), 4.24 (br d, J=13.4 Hz, 2H), 3.31 - 3.25 (m, IH), 3.02 (br s, 2H), 2.15 (br d, J=11.5 Hz, 2H), 1.76 (dq, J=4.2, 12.3 Hz, 2H), 1.28 (d, J=6.2 Hz, 6H), 1.11 (s, 9H). ESI [M+H] = 615.2
Example 5. Synthesis of isopropyl 4-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonyl amino)phenyl]thiazol-2-yl]piperazine-l-carboxylate.
Scheme 4:
Figure imgf000062_0002
Figure imgf000063_0001
Figure imgf000063_0002
General method A, tert-butyl 4-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl] thiazol-2- yl]piperazine-l-carboxylate. ESI [M+H] = 538.2
General method Cfor preparation of sulfonamide compound 57.
Figure imgf000063_0003
56
To a solution of tert-butyl 4-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl]thiazol -2- yl]piperazine-l-carboxylate (110 mg, 205 umol, 1 eq.) in DCM (2 mL) was added TFA (1 mL) and the mixture was stirred at 25°C for 30 mins. The mixture was concentrated to give N-[3-(tert-butylsulfamoyl)-4-(2-piperazin-l- ylthiazol-5-yl)phenyl]acetamide (80 mg, 183 umol, 89.4% yield) as a yellow solid. ESI [M+H] = 438.2 Preparation of compound 58
Figure imgf000064_0001
General method D, isopropyl 4-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl] thiazol-2- yljpiperazine-l-carboxylate. ESI [M+H] = 524.1
Preparation of compound 59
Figure imgf000064_0002
58
General method F, isopropyl 4-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol -2- yljpiperazine-l-carboxylate. ESI [M+H] = 482.1
Preparation of compound Ex. 5
Figure imgf000064_0003
59 Ex. 5
General method D, isopropyl 4-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonyl amino)phenyl]thiazol-2-yl]piperazine-l-carboxylate. 1H NMR (400MHz, METHANOL-d4) δ = 8.22 (d, J=2.1 Hz, IH), 7.56 (dd, J=2.2, 8.3 Hz, IH), 7.30 (d, J=8.3 Hz, IH), 7.20 (s, IH), 4.94 - 4.81 (m, 2H), 3.61 - 3.43 (m, 8H), 1.20 (dd, J=6.2, 14.5 Hz, 12H), 1.07 (s, 9H). ESI [M+H] = 568.3 Example 6. Synthesis of isopropyl 4-[5-[4-(benzyloxycarbonylamino )-2-(tert- butylsulfamoyl)phenyl]thiazol-2-yl]piperazine-l-carboxylate
Scheme 5 :
Preparation of compound Ex. 6
Figure imgf000065_0001
59 Ex. 6
General method D, isopropyl 4-[5-[4-(benzyloxycarbonylamino)-2-(tert- butylsulfamoyl)phenyl]thiazol-2-yl]piperazine-l-carboxylate. 1H NMR (400MHz, METHANOL-d4) δ = 8.33 (d, J=2.0 Hz, 1H), 7.68 (dd, J=2.2, 8.4 Hz, 1H), 7.45 - 7.27 (m, 7H), 5.21 (s, 2H), 4.94 - 4.88 (m, 1H), 3.68 - 3.51 (m, 8H), 1.27 (d, J=6.4 Hz, 6H), 1.19 - 1.11 (m, 9H). ESI [M+H] = 616.3
Example 7. Synthesis of trans-isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[4- (cyclopentoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate.
Scheme 6:
Figure imgf000065_0002
84 Ex. 7 General method B for preparation of sulfonamide compound 83.
dioxane/H20, 80°C
Figure imgf000066_0001
82 83
To a mixture of tert-butyl (trans-4-(5-bromothiazol-2-yl)cyclohexyl)carbamate (1.3 g, 3.6 mmol, 2 eq.) and isopropyl N-[3-(tert-butylsulfamoyl)-4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)phenyl]carbamate (800 mg, 1.8 mmol, 1 eq.) in dioxane (12 mL) and H20 (2 mL) were added Na2C03 (579 mg, 5.5 mmol, 3 eq.) and Pd(dppf)Cl2 (133 mg, 182 umol, 0.1 eq.). The mixture was stirred at 80°C for 12 hrs under N2 atmosphere and then concentrated. The residue was diluted with H20 (5 mL) and extracted with ethyl acetate (5 mL x 3), dried over Na2S04, filtered and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether: Ethyl acetate = 1 :0 to 3: 1) to give trans-isopropyl- N-[4-[2-[4-(tert-butoxycarbonylamino)cyclohexyl] thiazol-5-yl]-3- (tert- butylsulfamoyl)pheny I] carbamate (660 mg, crude), in which 60 mg was purified by prep- HPLC (column: Agela Durashell C18 150X25 5u;mobile phase: [water(0.04%NH3H2O)- ACN] ;B% : 60%-90%, 10min) to give pure compound 83 (40.92 mg, 99.64% purity) as a white solid for delivery. 1H NMR (400MHz, DMSO-d6) δ = 10.06 (s, IH), 8.32 (d, J=2.1 Hz, IH), 7.70 - 7.60 (m, 2H), 7.38 (d, J=8.4 Hz, IH), 6.96 (s, IH), 6.81 (br d, J=7.9 Hz, IH), 4.99 - 4.87 (m, IH), 3.27 (br s, IH), 2.94 - 2.84 (m, IH), 2.13 (br d, J=11.7 Hz, 2H), 1.90 (br d, J=l l . l Hz, 2H), 1.63 - 1.49 (m, 2H), 1.39 (s, 9H), 1.33 (br d, J=14.4 Hz, 2H), 1.28 (d, J=6.4 Hz, 6H), 1.07 (s, 9H). ESI [M+H] =595.3
Preparation of compound 84
Figure imgf000066_0002
83 84
General method F, trans-isopropyl N-[4-[2-(4-aminocyclohexyl)thiazol-5-yl]-3- (tert- butylsulfamoyl)pheny I] carbamate. ESI [M+H] = 495.2
Figure imgf000067_0001
General method D, trans-isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[4- (cyclopentoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate. 1H NMR
(400MHz, DMSO-d6) δ = 10.04 (s, IH), 8.29 (d, J=2.2 Hz, IH), 7.61 - 7.59 (m, 2H),7.35 (d, J=8.8 Hz, IH), 7.00 (br d, J=7.9 Hz, IH), 6.96 - 6.93 (m, IH), 4.96 - 4.87 (m, 2H), 3.29 - 3.22 (m, IH), 2.87 (br t, J=11.8 Hz, IH), 2.10 (br d, J=12.7 Hz, 2H), 1.88 (br d, J=12.7 Hz, 2H), 1.75 (br s, 2H), 1.61 - 1.48 (m, 8H), 1.36 - 1.27 (m, 2H), 1.25 (d, J=6.1 Hz, 6H), 1.04 (s, 9H). ESI [M+Na] = 629.3
Example 8. Synthesis of [(2R)-l-methylpyrrolidin-2-yl]methyl N-[4-[5-[2-(tert- butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate. Scheme 7:
Figure imgf000067_0002
Ex. 8
Preparation of compound 85
Figure imgf000067_0003
General method D, trans-(4-nitrophenyl) N-[4-[5-[2-(tert-butylsulfamoyl)-4- (isopropoxycarbonylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate. ESI [M+H]
Preparation of compound Ex. 8
Figure imgf000068_0001
General method H, [(2R)-l-methylpyrrolidin-2-yl]methyl N-[4-[5-[2-(tert- butylsulfamoyl)- 4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate. 1H NMR
(400MHz, METHANOL-d4) δ = 8.38 (s, IH), 7.80 - 7.73 (m, IH), 7.69 (dd, J=2.2, 8.4 Hz, IH), 7.40 (d, J=8.3 Hz, IH), 5.05 - 5.00 (m, IH), 4.48 (br dd, J=3.0, 12.8 Hz, IH), 4.21 (br dd, J=7.2, 12.6 Hz, IH), 3.78 - 3.66 (m, 2H), 3.57 - 3.46 (m, IH), 3.22 (td, J=8.3, 11.2 Hz, IH), 3.07 - 3.01 (m, 3H), 2.43 - 1.99 (m, 8H), 1.98 - 1.88 (m, IH), 1.80 - 1.66 (m, 2H), 1.47 (q, J=12.5 Hz, 2H), 1.34 (d, J=6.2 Hz, 6H), 1.20 - 1.12 (m, 9H). ESI [M+H] = 636.2
Example 9. Synthesis of isopropyl (3-(N-(tert-butyl)sulfamoyl)-4-(2-((lS,4r)-4-(((((S)- l-methylpyrrolidin-2-yl)methoxy)carbonyl)amino)cyclohexyl)thiazol-5- yl)phenyl)carbamate.
The following compound was synthesized via same method by the key intermediate 85.
Figure imgf000068_0002
Ex. 9
1H NMR (METHANOL-d4, 400MHz): δ = 8.34 (d, J=2.2 Hz, IH), 7.71 (s, IH), 7.66 (dd, J=8.5, 2.1 Hz, IH), 7.36 (d, J=8.4 Hz, IH), 4.93-5.04 (m, IH), 4.46 (dd, J=13.0, 2.9 Hz, IH), 4.18 (dd, J=13.0, 7.1 Hz, IH), 3.64-3.75 (m, 2H), 3.42-3.58 (m, IH), 3.12-3.25 (m, IH), 3.00- 3.06 (m, 3H), 1.97-2.39 (m, 8H), 1.84-1.96 (m, IH), 1.64-1.77 (m, 2H), 1.44 (q, J=12.6 Hz, 2H), 1.31 (d, J=6.2 Hz, 6H), 1.13 (s, 9H). ESI [M+H] = 636.3 Example 10. Synthesis of trans-tert-butyl N-[4-[5-[4-(benzylcarbamoylamino)-2- (tert- butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate.
Figure imgf000069_0001
General method B, trans-tert-butyl N-[4-[5-[4-(benzylcarbamoylamino)-2- (tert- butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate. 1H NMR (400MHz,
METHANOL-d4) δ = 8.24 (d, J=2.4 Hz, IH), 7.72 - 7.66 (m, 2H), 7.37 - 7.29 (m, 5H), 7.28 - 7.21 (m, IH), 4.40 (s, 2H), 3.39 (br t, J=11.6 Hz, IH), 2.98 (tt, J=3.4, 12.1 Hz, IH), 2.25 - 2.17 (m, 2H), 2.04 (br d, J=11.5 Hz, 2H), 1.67 (dq, J=2.9, 12.9 Hz, 2H), 1.44 (s, 9H), 1.41 - 1.32 (m, 2H), 1.10 (s, 9H). ESI [M+H] = 642.3
Example 11. Synthesis of [(2R)-l-methylpyrrolidin-2-yl]methyl N-[4-[5-[4- (benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbam
Scheme 8:
Figure imgf000069_0002
Ex. 11 Preparation of compound 86
Figure imgf000070_0001
Ex. 10 86
General method F, trans-l-[4-[2-(4-aminocyclohexyl)thiazol-5-yl]-3-(tert- butylsulfamoyl)phenyl]-3-benzyl-urea. ESI [M+H] = 542.3
Preparation of compound 87
Figure imgf000070_0002
General method D, trans-(4-nitrophenyl) N-[4-[5-[4-(benzylcarbamoylamino)-2- (tert- butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate.
Preparation of Ex. 11
Figure imgf000070_0003
General method H, [(2R)-l-methylpyrrolidin-2-yl]methyl N-[4-[5-[4- (benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate.
1H NMR (400MHz, METHANOL-d4) δ = 8.30 (d, J=2.2 Hz, 1H), 7.79 (s, 1H), 7.70 (dd, J=2.3, 8.4 Hz, 1H), 7.42 - 7.31 (m, 5H), 7.27 (dt, J=2.6, 5.7 Hz, 1H), 4.54 - 4.41 (m, 3H), 4.21 (dd, J=7.1, 12.7 Hz, 1H), 3.79 - 3.66 (m, 2H), 3.52 (br t, J=11.6 Hz, 1H), 3.26 - 3.17 (m, 1H), 3.04 (s, 3H), 2.42 - 1.98 (m, 8H), 1.97 - 1.87 (m, 1H), 1.80 - 1.65 (m, 2H), 1.54 - 1.40 (m, 2H), 1.15 (s, 9H). ESI [M+H] = 683.3 Example 12. Synthesis of ((S)-l-methylpyrrolidin-2-yl)methyl ((lr,4S)-4-(5-(4-(3- benzylureido)-2-(N-(tert-butyl)sulfamoyl)phenyl)thiazol-2-yl)cyclohexyl)carbamate
The following compound was synthesized via same method by the key intermediate 87.
Figure imgf000071_0001
Ex. 12
1H NMR (400MHz, METHANOL-d4) δ = 8.29 (d, J=2.3 Hz, 1H), 7.83 - 7.74 (m, 1H), 7.73 - 7.65 (m, 1H), 7.43 - 7.31 (m, 5H), 7.30 - 7.22 (m, 1H), 4.55 - 4.35 (m, 3H), 4.21 (br dd, J=7.1, 12.7 Hz, 1H), 3.79 - 3.61 (m, 2H), 3.58 - 3.42 (m, 1H), 3.23 (br s, 1H), 3.08 - 3.02 (m, 3H), 2.43 - 1.98 (m, 8H), 1.97 - 1.86 (m, 1H), 1.80 - 1.65 (m, 2H), 1.54 - 1.39 (m, 2H), 1.19 - 1.07 (m, 9H). ESI [M+H] = 683.3
Example 13. Synthesis of isopropyl (S)-(4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(l- phenylethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-l-yl)carbamate.
Scheme 9:
Figure imgf000071_0002
Figure imgf000072_0001
Ex. 13
Preparation of compound 89
Figure imgf000072_0002
88 89
General method D, 4-(isopropoxycarbonylamino)bicyclo[2.2.2]octane-l-carboxylate. General method O for preparation of compound 90
Figure imgf000072_0003
89 90
To a solution of methyl 4-(isopropoxycarbonylamino)bicyclo[2.2.2]octane-l- carboxylate (400 mg, 1.49 mmol, 1 eq.) in MeOH (5 mL), was added LiOH (106.70 mg, 4.46 mmol, 3 eq.) in H20 (5 mL) and the mixture was stirred at 50°C for 1 hr. The mixture was concentrated to remove MeOH and then extracted with MTBE (10 mL*2). The pH of aqueous phase was adjusted to 1-2 by adding 4 N HC1 solution and then extracted with EtOAc (10 mL*3). The combined organic phase was dried over Na2S04, filtered and concentrated to give 4-(isopropoxycarbonylamino)bicyclo[2.2.2]octane-l-carboxylic acid (270 mg, crude) as a yellow solid. 1H NMR (400MHz, CHLOROFORM-d) δ = 4.86 (br d, J=6.4 Hz, 1H), 4.46 (br s, 1H), 2.00 - 1.80 (m, 12H), 1.23 (br d, J=5.9 Hz, 6H).
General method N for preparation of compound 91
Figure imgf000072_0004
90 91 To a solution of 4-(isopropoxycarbonylamino)bicyclo[2.2.2]octane-l-carboxylic acid (270 mg, 1.06 mmol, 1 eq.) in DCM (5 mL), were added DMF (7.73 mg, 105.75 umol, 0.1 eq.) and (COCl)2 (201.34 mg, 1.59 mmol, 1.5 eq.). The mixture was stirred at 26°C for 0.2 hr and then concentrated. The residue was dissolved into THF (5 mL) and was added into NH3.H20 (741.24 mg, 5.29 mmol, 5 eq.) in dioxane (5 mL) dropwise. Then the mixture was stirred at 26°C for 0.3 hr. The mixture was concentrated and diluted with EtOAc (20 mL) and washed with H20 (10 mL). The organic phase was dried over Na2S04, fitlered and concentrated to give isopropyl N-(l-carbamoyl-4-bicyclo[2.2.2]octanyl)carbamate as a yellow solid. 1H NMR (400MHz, CHLOROFORM-d) δ = 5.43 (br s, 1H), 5.17 (br s, 1H), 4.85 - 4.71 (m, 1H), 4.35 (br s, 1H), 1.82 (s, 12H), 1.13 (d, J=6.2 Hz, 6H). ESI[M+H]=255.2
General method Lfor preparation of compound 92
Figure imgf000073_0001
91
To a solution of isopropyl N-(l-carbamoyl-4-bicyclo[2.2.2]octanyl)carbamate (60 mg,
235.92 umol, 1 eq.) in 2-Me-THF (2 mL), was added 2,4-bis(4-methoxyphenyl)-2,4-dithioxo- l,3,2,4dithiadiphosphetane (95.42 mg, 235.92 umol, 1 eq.) and the mixture was stirred at 80°C for 0.5 hr. The mixture was poured into sat.aq.Na2C03 (10 mL) and extracted with EtOAc (10 mL*3). The combined organic phase was dried over Na2S04, filtered and concentrated. The residue was purified by prep-TLC (Petroleum ether/Ethyl acetate =2:3) to give isopropyl N-(l-carbamothioyl-4-bicyclo[2.2.2]octanyl)carbamate (30 mg, crude) as a yellow solid. ESI [M+H]=271.1
General method Mfor preparation of compound 93
Figure imgf000073_0002
92 93
To a solution of isopropyl N-(l-carbamothioyl-4-bicyclo[2.2.2]octanyl)carbamate (85 mg,
314.36 umol, 1 eq.) in EtOH (2 mL), were added TsOH.H20 (119.59 mg, 628.72 umol, 2 eq.) and 2-bromo- l, l-diethoxy-ethane (123.90 mg, 628.72 umol, 2 eq.). The mixture was stirred at 80°C for 1 hr and then concentrated and diluted with EtOAc (30 mL). The mixture was washed with sat.aq.Na2C03 (10 mL*2) and the combined organic phase was dried over Na2S04, filtered and concentrated. The residue was purified by prep-TLC (Petroleum ether/Ethyl acetate = 3: 1) to give isopropyl N-(l-thiazol-2-yl-4- bicyclo[2.2.2]octanyl)carbamate (80 mg, crude) as a yellow solid. ESI [M+H]=295.3
Preparation of compound 94
Figure imgf000074_0001
93 94
General method J, isopropyl N-[l-(5-bromothiazol-2-yl)-4-bicyclo[2.2.2] octanyljcarbamate. ESI[M+H]=375.2/373.2
Preparation of compound 95
Figure imgf000074_0002
General method B, isopropyl N-[l-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl] thiazol-2-yl]- 4-bicyclo[2.2.2]octanyl]carbamate. ESI [M+H] = 521.2
Preparation of compound 96
Figure imgf000074_0003
General method D, (4-nitrophenyl) N-[3-(tert-butylsulfamoyl)-4-[2-[4- (isopropoxycarbonylamino)-l-bicyclo[2.2.2]octanyl]thiazol-5-yl]phenyl]carbamate. ESI [M+H] = 686.4 Preparation of Ex. 13
Figure imgf000075_0001
General method E, isopropyl N-[l-[5-[2-(tert-butylsulfamoyl)-4-[[(lS)-l-phenylethyl] carbamoylamino ]phenyl]thiazol-2-yl]-4-bicyclo[2.2.2]octanyl]carbamate. 1H NMR
(400MHz, METHANOL-d4) δ = 8.21 (d, J=2.3 Hz, 1H), 7.75 (s, 1H), 7.67 (dd, J=2.3, 8.3 Hz, 1H), 7.41 - 7.32 (m, 5H), 7.29 - 7.23 (m, 1H), 4.99 - 4.93 (m, 1H), 4.83 - 4.76 (m, 1H), 2.17 - 2.08 (m, 6H), 2.07 - 1.99 (m, 6H), 1.51 (d, J=7.0 Hz, 3H), 1.22 (br d, J=6.0 Hz, 6H), 1.12 (s, 9H). ESI [M+H] = 668.3
Example 14. Synthesis of isopropyl (R)-(4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(l- phenylethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-l-yl)carbamate.
The following compound was synthesized via same method by the key intermediate 96.
Figure imgf000075_0002
Ex. 14
1H NMR (400MHz, METHANOL-d4) δ = 8.18 (d, J=2.6 Hz, 1H), 7.74 - 7.69 (m, 1H), 7.64 (dd, J=2.2, 8.3 Hz, 1H), 7.38 - 7.29 (m, 5H), 7.26 - 7.19 (m, 1H), 4.92 (q, J=7.0 Hz, 1H), 4.82 - 4.71 (m, 1H), 2.15 - 2.05 (m, 6H), 2.04 - 1.95 (m, 6H), 1.48 (d, J=7.0 Hz, 3H), 1.20 (br d, J=6.1 Hz, 6H), 1.09 (s, 9H). ESI [M+H] = 668.3
Example 15. Synthesis of isopropyl (S)-(4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(l- (pyridin-2-yl)ethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-l-yl)carbamate.
The following compound was synthesized via same method by the key intermediate 96.
Figure imgf000075_0003
Ex. 15 1H NMR (400MHz, METHANOL-d4) δ = 8.72 (d, J=5.6 Hz, IH), 8.46 (t, J=7.9 Hz, IH), 8.28 (d, J=2.3 Hz, IH), 8.02 (d, J=7.9 Hz, IH), 7.85 (t, J=6.6 Hz, IH), 7.72 (s, IH), 7.63 (dd, J=2.3, 8.4 Hz, IH), 7.35 (d, J=8.4 Hz, IH), 5.13 (q, J=7.1 Hz, IH), 4.84 - 4.74 (m, IH), 2.15 - 2.07 (m, 6H), 2.07 - 1.98 (m, 6H), 1.65 (d, J=7.1 Hz, 3H), 1.22 (br d, J=6.1 Hz, 6H), 1.09 (s, 9H). ESI [M+H] = 669.2
Example 16. Synthesis of isopropyl (R)-(4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(l- (pyridin-2-yl)ethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-l-yl)carbamate.
The following compound was synthesized via same method by the key intermediate 96.
Figure imgf000076_0001
Ex. 16
1H NMR (400MHz, METHANOL-d4) δ = 8.75 (d, J=5.1 Hz, IH), 8.57 (dt, J=1.5, 7.9 Hz, IH), 8.29 (d, J=2.2 Hz, IH), 8.11 (d, J=8.1 Hz, IH), 7.98 - 7.90 (m, IH), 7.75 (s, IH), 7.62 (dd, J=2.3, 8.3 Hz, IH), 7.36 (d, J=8.4 Hz, IH), 5.15 (q, J=7.1 Hz, IH), 4.84 - 4.75 (m, IH), 2.16 - 2.07 (m, 6H), 2.06 - 1.97 (m, 6H), 1.67 (d, J=7.2 Hz, 3H), 1.22 (br d, J=6.1 Hz, 6H), 1.09 (s, 9H). ESI [M+H] = 669.2
Example 17. Synthesis of isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[4- (isopropoxycarbonyl amino)-l-bicyclo[2.2.2]octanyl]thiazol-5-yl]phenyl]carbamate.
Scheme 10:
Preparation of compound Ex. 17
Figure imgf000076_0002
Ex. 17
General method B, isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonyl amino)-l-bicyclo[2.2.2]octanyl]thiazol-5-yl]phenyl]carbamate. 1H NMR (400MHz, METHANOL-d4) δ = 8.33 (d, J=2.0 Hz, IH), 7.74 (s, IH), 7.66 (dd, J=2.2, 8.4 Hz, IH), 7.36 (d, J=8.4 Hz, IH), 5.03 - 4.91 (m, IH), 4.78 (br d, J=6.4 Hz, IH), 2.16 - 2.05 (m, 6H), 2.04 - 1.95 (m, 6H), 1.31 (d, J=6.4 Hz, 6H), 1.20 (br d, J=6.0 Hz, 6H), 1.11 (s, 9H). ESI [M+H] =607.3
Example 18. Synthesis of isopropyl (4-(5-(4-(3-benzylureido)-2-(N-(tert- butyl)sulfamoyl)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-l-yl)carbamate.
The following compound was synthesized via same method by the key intermediate 94.
Figure imgf000077_0001
Ex. 18
1H NMR (400MHz, METHANOL-d4) δ = 8.26 (d, J=2.2 Hz, IH), 7.77 (s, IH), 7.72 (dd, J=2.4, 8.4 Hz, IH), 7.38 - 7.33 (m, 5H), 7.27 (td, J=2.6, 8.6 Hz, IH), 4.80 (br d, J=5.7 Hz, IH), 4.43 (s, 2H), 2.17 - 2.10 (m, 6H), 2.06 - 2.00 (m, 6H), 1.25 - 1.20 (m, 6H), 1.13 (s, 9H). ESI [M+H] =654.1
Example 19. Synthesis of isopropyl (4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(pyridin-2- ylmethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-l-yl)carbamate.
The following compound was synthesized via same method by the key intermediate 94.
Figure imgf000077_0002
Ex. 19
1H NMR (400MHz, METHANOL-d4) δ = 8.73 (d, J=5.7 Hz, IH), 8.51 (dt, J=1.3, 7.9 Hz, IH), 8.31 (d, J=2.2 Hz, IH), 8.02 (d, J=8.3 Hz, IH), 7.90 (t, J=6.8 Hz, IH), 7.72 (s, IH), 7.66 (dd, J=2.2, 8.3 Hz, IH), 7.35 (d, J=8.3 Hz, IH), 4.75 (s, 3H), 2.15 - 1.93 (m, 12H), 1.20 (br d, J=6.1 Hz, 6H), 1.07 (s, 9H). ESI [M+H] =655.3 Example 20. Synthesis of trans-isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[4- ( isopropylcarbamoyloxy )cyclohexyl]thiazol-5-yl]phenyl]carbamate .
Scheme 11 :
OTBDPS
TEA, MeCN imidazole, DCM
Figure imgf000078_0001
110
Lawessons Reagent NBS
' OTBDPS
Na2C03, THF
Figure imgf000078_0002
DMF, 25°C
1 13 114
Figure imgf000078_0003
114A 115 116
Figure imgf000078_0004
Ex. 20
Preparation of compound 111.
Figure imgf000078_0005
110 11 1
To a solution of trans-4-hydroxycyclohexanecarboxylic acid (1 g, 6.94 mmol, 1 eq.) in MeCN (10 mL), were added HBTU (2.89 g, 7.63 mmol, 1.1 eq.) and TEA (2.11 g, 20.81 mmol, 2.90 mL, 3 eq.) followed by addition of NH4C1 (742.07 mg, 13.87 mmol, 2 eq.) and the mixture was stirred at 26°C for 1 hr. The mixture was then filtered and dried to give trans-4-hydroxycyclohexanecarboxamide (1 g, crude) as a white solid which can be used without any purification.
Preparation of compound 112.
OTBDPS
imidazole, DCM
Figure imgf000078_0006
111 112 To a solution of trans-4-hydroxycyclohexanecarboxamide (12 g, 83.81 mmol, 1 eq.) in DCM (150 mL), were added IMIDAZOLE (11.41 g, 167.62 mmol, 2 eq.) and tert-butyl- chloro-diphenyl-silane (27.64 g, 100.57 mmol, 25.83 mL, 1.2 eq.). The mixture was stirred at 26°C for 12 hrs and then poured into IN HCl (200 mL) and the organic phase was washed with sat.aq.Na2C03 (100 mL). The organic phase was dried over Na2S04, filtered and concentrated. The residue was washed with a solution (petroleum ether : EtOAc=10: l, 100 mL) and then filtered. The filter cake was dried to give trans4-[tert- butyl(diphenyl)silyl]oxycyclohexanecarboxamide (31.5 g, crude). 1H NMR (400MHz, METHANOL-d4) δ = 7.72 - 7.64 (m, 4H), 7.49 - 7.36 (m, 6H), 3.69 - 3.55 (m, 1H), 2.21 - 2.08 (m, 1H), 1.96 - 1.83 (m, 2H), 1.82 - 1.69 (m, 2H), 1.50 - 1.23 (m, 4H), 1.12 - 0.98 (m, 9H).
Preparation of compound 113.
OTBDPS
Figure imgf000079_0001
112
General method L, trans-4-[tert-butyl(diphenyl)silyl]oxycyclohexanecarbothioamide. 1H
NMR (400MHz, METHANOL-d4) δ = 7.73 - 7.64 (m, 4H), 7.52 - 7.36 (m, 6H), 3.73 - 3.59 (m, 1H), 2.59 - 2.44 (m, 1H), 1.91 (br d, J=8.3 Hz, 2H), 1.80 - 1.66 (m, 2H), 1.57 - 1.35 (m, 4H), 1.08 - 1.04 (m, 9H). ESI [M+H] =398.1
Preparation of compound 114. " 'OTBDPS
Figure imgf000079_0002
113 114
General method M, trans-tert-butyl-diphenyl-(4-thiazol-2-ylcyclohexoxy)silane. 1H NMR
(400MHz, METHANOL-d4) δ = 7.63 - 7.55 (m, 5H), 7.42 - 7.25 (m, 7H), 4.05 - 3.96 (m, 1H), 2.97 (tt, J=3.7, 11.7 Hz, 1H), 2.05 (dq, J=3.2, 12.5 Hz, 2H), 1.84 - 1.62 (m, 4H), 1.44 (tt, J=2.9, 13.5 Hz, 2H), 1.01 - 0.98 (m, 9H). ESI [M+H] =422.2 Preparation of compound 114 A.
Figure imgf000080_0001
General method J, trans-[4-(5-bromothiazol-2-yl)cyclohexoxy]-tert-butyl-diphenyl -silane (1.8 g, crude). ESI [M+H] =502.0
Preparation of compound 115.
Figure imgf000080_0002
To a solution of trans-[4-(5-bromothiazol-2-yl)cyclohexoxy]-tert-butyl-diphenyl -silane (1.5 g, 3.00 mmol, 1 eq.) in THF (10 niL), was added TBAF (1 M, 4.49 mL, 1.5 eq.) and the mixture was stirred at 26°C for 12 hrs and then concentrated. The residue was purified by silica gel chromatography (Petroleum ether/Ethyl acetate=20: 1- 1 : 1) to afford trans-4-(5- bromothiazol-2-yl)cyclohexanol (500 mg, 1.91 mmol, 63.64% yield) as a yellow solid.
Preparation of compound 116.
Figure imgf000080_0003
115 116
To a solution of trans-4-(5-bromothiazol-2-yl)cyclohexanol (500 mg, 1.91 mmol, 1 eq.) in DMF (5 mL), were added 2-isocyanatopropane (486.93 mg, 5.72 mmol, 3 eq.) and DIEA (739.47 mg, 5.72 mmol, 3 eq.). The mixture was stirred at 100°C for 40 hrs and then poured into H20 (50 mL) and extracted with EtOAc (10 mL*3). The combined organic phase was dried over Na2S04, filtered and concentrated. The residue was purified by silica gel chromatography (Petroleum ether/Ethyl acetate=10: 1- 1 : 1) to afford trans-[4-(5- bromothiazol-2-yl)cyclohexyl] N-isopropylcarbamate (180 mg, 518.33 umol, 27.18% yield) as a yellow solid. 1H NMR (400MHz, METHANOL-d4) δ = 7.61 (s, 1H), 3.85 - 3.64 (m, 2H), 3.15 - 2.99 (m, 1H), 1.98 - 1.83 (m, 6H), 1.73 (br d, J=13.5 Hz, 2H), 1.14 - 1.12 (m, 6H). ESI [M+H] =347.1/349.1 Preparation of Ex. 20.
Figure imgf000081_0001
dioxane/H20, 80°C
116 Ex. 20
General method B, trans-isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[4- (isopropylcarbamoyloxy)cyclohexyl]thiazol-5-yl]phenyl]carbamate. 1H NMR (400MHz, METHANOL-d4) δ = 8.34 (d, J=2.0 Hz, 1H), 7.75 (s, 1H), 7.67 (dd, J=2.0, 8.4 Hz, 1H), 7.37 (d, J=8.4 Hz, 1H), 5.05 - 4.93 (m, 1H), 4.89 (br s, 1H), 3.83 - 3.64 (m, 1H), 3.15 (br s, 1H), 2.12 - 1.88 (m, 6H), 1.84 - 1.64 (m, 2H), 1.31 (d, J=6.2 Hz, 6H), 1.19 - 1.04 (m, 15H). ESI [M+H] =581.4
Example 21. Synthesis of (lr,4r)-4-(5-(4-(3-benzylureido)-2-(N-(tert-butyl)sulfamoyl) phenyl)thiazol-2-yl)cyclohexyl isopropylcarbamate
The following compound was synthesized via same method by the key intermediate 116.
Figure imgf000081_0002
1H NMR (400MHz, METHANOL-d4) δ = 8.24 (d, J=2.2 Hz, 1H), 7.74 (s, 1H), 7.69 (dd, J=2.4, 8.4 Hz, 1H), 7.40 - 7.28 (m, 5H), 7.28 - 7.17 (m, 1H), 4.89 (br s, 1H), 4.41 (s, 2H), 3.79 - 3.64 (m, 1H), 3.14 (br s, 1H), 2.04 - 1.94 (m, 6H), 1.84 - 1.65 (m, 2H), 1.16 - 1.08 (m, 15H). ESI [M+H] = 628.4
Example 22. Synthesis of trans-isopropyl N-[4-[5-[2-(tert-butylsulfamoyl)-4-(4 - pyridylmethylcarbamoylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate.
Scheme 12:
Figure imgf000081_0003
Figure imgf000082_0001
Preparation of compound 117.
Figure imgf000082_0002
General method K, trans-isopropyl N-[4-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl] thiazol-2-yl]cyclohexyl]carbamate. IH NMR (400MHz, METHANOL-d4) δ = 7.68 - 7.57 (s, IH), 7.43 (d, J=2.4 Hz, IH), 7.13 (d, J=8.2 Hz, IH), 6.82 (dd, J=2.4, 8.2 Hz, IH), 4.84 - 4.77 (m, IH), 3.44 (tt, J=3.9, 11.5 Hz, IH), 2.97 (tt, J=3.6, 12.1 Hz, IH), 2.26 - 2.13 (m, 2H), 2.11 - 2.02 (m, 2H), 1.67 (dq, J=3.0, 12.9 Hz, 2H), 1.49 - 1.32 (m, 2H), 1.22 (dd, J=2.5, 6.7 Hz, 6H), 1.09 (s, 9H). ESI [M+H] =495.2
Preparation of compoundll8.
Figure imgf000082_0003
117 118
General method D, trans-(4-nitrophenyl) N-[3-(tert-butylsulfamoyl)-4-[2-[4- (isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate. ESI [M+H] =660.1 Preparation of Ex. 22.
Figure imgf000083_0001
General method E, trans-isopropyl N-[4-[5-[2-(tert-butylsulfamoyl)-4-(4 - pyridylmethylcarbamoylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate. 1H NMR
(400MHz, METHANOL-d4) δ = 8.79 (d, J=6.7 Hz, 2H), 8.34 (d, J=2.3 Hz, 1H), 8.07 (d, J=6.6 Hz, 2H), 7.76 (s, 1H), 7.69 (dd, J=2.3, 8.4 Hz, 1H), 7.39 (d, J=8.3 Hz, 1H), 4.85 - 4.83 (m, 1H), 4.73 (s, 2H), 3.47 (tt, J=3.6, 11.5 Hz, 1H), 3.04 (tt, J=3.4, 12.0 Hz, 1H), 2.30 - 2.19 (m, 2H), 2.09 (br d, J=10.1 Hz, 2H), 1.71 (dq, J=3.0, 12.9 Hz, 2H), 1.43 (dq, J=3.3, 12.6 Hz, 2H), 1.24 (br d, J=6.1 Hz, 6H), 1.12 (s, 9H). ESI [M+H] =629.2
Example 23. Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(4- fluorobenzyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000083_0002
Ex. 23
1H NMR (400MHz, METHANOL-d4) δ = 8.27 (d, J=2.3 Hz, 1H), 7.77 (s, 1H), 7.71 (dd, J=2.4, 8.4 Hz, 1H), 7.38 (td, J=2.7, 8.6 Hz, 3H), 7.12 - 7.04 (m, 2H), 5.00 - 4.92 (m, 1H), 4.41 (s, 2H), 3.54 - 3.41 (m, 1H), 3.05 (tt, J=3.5, 12.0 Hz, 1H),2.13 - 2.05 (m, 2H), 2.05 - 2.04 (m, 2H), 1.72 (dq, J=3.0, 12.8 Hz, 2H), 1.43 (dq, J=3.2, 12.5 Hz, 2H), 1.24 (br d, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H] =646.2 Example 24. Synthesis of isopropyl ((lR,4r)-4-(5-(2-(N-isopropylsulfamoyl)-4-(3-((R)-l- phenylethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000084_0001
Ex. 24
1H NMR (400MHz, METHANOL-d4) δ = 8.20 (d, J=2.2 Hz, IH), 7.73 (s, IH), 7.64 (dd, J=2.4, 8.4 Hz, IH), 7.38 - 7.29 (m, 5H), 7.26 - 7.19 (m, IH), 4.94 - 4.89 (m, IH), 4.81 (br d, J=6.2 Hz, IH), 3.44 (tt, J=3.9, 11.6 Hz, IH), 3.01 (tt, J=3.5, 12.0 Hz, IH), 2.26 - 2.17 (m, 2H), 2.10 - 2.01 (m, 2H), 1.68 (dq, J=2.9, 12.8 Hz, 2H), 1.48 (d, J=7.1 Hz, 3H), 1.43 - 1.33 (m, 2H), 1.21 (br d, J=6.2 Hz, 6H), 1.10 (s, 9H). ESI [M+H] =642.3
Example 25. Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3- (pyridin-3-ylmethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000084_0002
Ex. 25
1H NMR (400MHz, METHANOL-d4) δ = 8.84 (s, IH), 8.74 (d, J=5.7 Hz, IH), 8.60 (d, J=8.2 Hz, IH), 8.30 (d, J=2.2 Hz, IH), 8.04 (dd, J=5.8, 7.8 Hz, IH), 7.72 (s, IH), 7.65 (dd, J=2.4, 8.4 Hz, IH), 7.36 (d, J=8.4 Hz, IH), 4.85 - 4.77 (m, IH), 4.60 (s, 2H), 3.44 (tt, J=3.9, 11.5 Hz, IH), 3.10 - 2.93 (m, IH), 2.28 - 2.17 (m, 2H), 2.13 - 2.01 (m, 2H), 1.69 (dq, J=3.0, 12.8 Hz, 2H), 1.50 - 1.33 (m, 2H), 1.22 (br d, J=6.2 Hz, 6H), 1.09 (s, 9H). ESI [M+H] =629.3 Example 26. Synthesis of isopropyl ((lS,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((S)- l-phenylethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000085_0001
Ex. 26
1H NMR (400MHz, METHANOL-d4) δ = 8.19 (d, J=2.4 Hz, IH), 7.70 (s, IH), 7.65 (dd, J=2.4, 8.4 Hz, IH), 7.39 - 7.30 (m, 5H), 7.26 - 7.21 (m, IH), 4.93 (q, J=6.5 Hz, IH), 4.83 - 4.81 (m, IH), 3.49 - 3.40 (m, IH), 3.05 - 2.96 (m, IH), 2.22 (br d, J=11.7 Hz, 2H), 2.06 (br d, J=11.5 Hz, 2H), 1.73 - 1.62 (m, 2H), 1.49 (d, J=6.8 Hz, 3H), 1.43 - 1.36 (m, 2H), 1.22 (br d, J=6.0 Hz, 6H), 1.10 (s, 9H). ESI[M+H] =642.3
Example 27. Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(3- fluorobenzyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000085_0002
Ex. 27
1H NMR (400MHz, METHANOL-d4) δ = 8.27 (d, J=2.2 Hz, IH), 7.74 (s, IH), 7.71 (dd, J=2.3, 8.3 Hz, IH), 7.40 - 7.32 (m, 2H), 7.18 (d, J=7.7 Hz, IH), 7.10 (br d, J=9.9 Hz, IH), 6.99 (dt, J=2.2, 8.5 Hz, IH), 4.87 (br s, IH), 4.44 (s, 2H), 3.47 (tt, J=3.8, 11.6 Hz, IH), 3.02 (tt, J=3.5, 12.0 Hz, IH), 2.24 (br d, J=12.2 Hz, 2H), 2.14 - 2.04 (m, 2H), 1.71 (dq, J=3.0, 12.9 Hz, 2H), 1.50 - 1.36 (m, 2H), 1.24 (br d, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H] =646.2 Example 28. Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(2- fluorobenzyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000086_0001
Ex. 28
1H NMR (400MHz, METHANOL-d4) δ = 8.23 (d, J=2.2 Hz, IH), 7.71 (s, IH), 7.68 (dd, J=2.4, 8.4 Hz, IH), 7.41 (dt, J=1.5, 7.6 Hz, IH), 7.35 (d, J=8.4 Hz, IH), 7.29 (ddt, J=1.8, 5.5, 7.7 Hz, IH), 7.15 (dt, J=1.0, 7.6 Hz, IH), 7.11 - 7.05 (m, IH), 4.81 (br s, IH), 4.46 (s, 2H), 3.45 (tt, J=3.7, 11.6 Hz, IH), 3.00 (tt, J=3.5, 12.0 Hz, IH), 2.27 - 2.18 (m, 2H), 2.06 (br d, J=10.1 Hz, 2H), 1.69 (dq, J=3.1, 12.9 Hz, 2H), 1.46 - 1.34 (m, 2H), 1.22 (br d, J=6.2 Hz, 6H), 1.11 (s, 9H). ESI [M+H] =646.2
Example 29. Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3- (pyridin-2-ylmethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000086_0002
Ex. 29
1H NMR (400MHz, METHANOL-d4) δ = 8.79 - 8.73 (m, IH), 8.56 (dt, J=1.5, 7.9 Hz, IH), 8.35 (d, J=2.2 Hz, IH), 8.06 (d, J=8.1 Hz, IH), 7.94 (t, J=6.4 Hz, IH), 7.77 (s, IH), 7.69 (dd, J=2.3, 8.4 Hz, IH), 7.39 (d, J=8.3 Hz, IH), 4.86 - 4.83 (m, IH), 4.79 (s, 2H), 3.47 (tt, J=3.9, 11.6 Hz, IH), 3.05 (tt, J=3.5, 12.0 Hz, IH), 2.29 - 2.21 (m, 2H), 2.09 (br d, J=10.1 Hz, 2H), 1.71 (dq, J=2.9, 12.8 Hz, 2H), 1.43 (dq, J=3.3, 12.6 Hz, 2H), 1.24 (br d, J=6.2 Hz, 6H), 1.11 (s, 9H). ESI [M+H] =629.3 Example 30. Synthesis of isopropyl ((lR,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((R)- l-(pyridin-2-yl)ethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000087_0001
Ex. 30
1H NMR (400MHz, METHANOL-d4) δ = 8.73 (d, J=5.7 Hz, 1H), 8.53 (dt, J=1.8, 7.9 Hz, 1H), 8.28 (d, J=2.2 Hz, 1H), 8.08 (d, J=8.3 Hz, 1H), 7.94 - 7.85 (m, 1H), 7.80 - 7.68 (m, 1H), 7.61 (dd, J=2.2, 8.3 Hz, 1H), 7.35 (d, J=8.3 Hz, 1H), 5.13 (q, J=7.0 Hz, 1H), 4.85 - 4.78 (m, 1H), 3.49 - 3.39 (m, 1H), 3.08 - 2.96 (m, 1H), 2.22 (br d, J=12.3 Hz, 2H), 2.06 (br d, J=10.1 Hz, 2H), 1.76 - 1.61 (m, 5H), 1.49 - 1.34 (m, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.14 - 1.02 (m, 9H). ESI [M+H] =643.3
Example 31. Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((3- fluoropyridin-2-yl)methyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000087_0002
Ex. 31
1H NMR (400MHz, METHANOL-d4) δ = 8.42 (d, J=4.8 Hz, 1H), 8.28 (d, J=2.2 Hz, 1H), 7.78 - 7.62 (m, 3H), 7.44 (td, J=4.4, 8.5 Hz, 1H), 7.38 (d, J=8.4 Hz, 1H), 4.86 - 4.81 (m, 1H), 4.65 (s, 2H), 3.47 (ddd, J=4.0, 7.7, 11.4 Hz, 1H), 3.10 - 3.00 (m, 1H), 2.25 (br d, J=12.1 Hz, 2H), 2.09 (br d, J=10.5 Hz, 2H), 1.72 (dq, J=3.0, 12.9 Hz, 2H), 1.43 (dq, J=3.2, 12.6 Hz, 2H), 1.24 (br d, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H] =647.2 Example 32. Synthesis of isopropyl ((lS,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((S)- l-(pyridin-2-yl)ethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000088_0001
1H NMR (400MHz, METHANOL-d4) δ = 8.72 (d, J=5.3 Hz, IH), 8.50 (dt, J=1.8, 7.9 Hz, IH), 8.27 (d, J=2.2 Hz, IH), 8.05 (d, J=8.3 Hz, IH), 7.88 (t, J=6.6 Hz, IH), 7.72 (s, IH), 7.61 (dd, J=2.6, 8.3 Hz, IH), 7.34 (d, J=8.8 Hz, IH), 5.12 (q, J=7.0 Hz, IH), 4.87 - 4.68 (m, IH), 3.51 - 3.38 (m, IH), 3.01 (tt, J=3.3, 12.0 Hz, IH), 2.21 (br d, J=11.8 Hz, 2H), 2.10 - 1.97 (m, 2H), 1.76 - 1.57 (m, 5H), 1.48 - 1.33 (m, 2H), 1.22 (d, J=6.1 Hz, 6H), 1.13 - 1.00 (m, 9H). ESI [M+H] =643.3
Example 33. Synthesis of trans-4-piperidylmethyl N-[3-(tert-butylsulfamoyl)-4-[2-[4- ( isopropoxycarbonylamino )cyclohexyl]thiazol-5-yl]phenyl]carbamate .
Scheme 13:
Figure imgf000088_0002
Ex. 33
Figure imgf000089_0001
General method H, trans-tert-butyl 4-[[3-(tert-butylsulfamoyl)-4-[2-[4- (isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbam
1-carboxylate. ESI [M+H] =736.5
Preparation of Ex. 33.
Figure imgf000089_0002
119 Ex. 33
General method C, trans-4-piperidylmethyl N-[3-(tert-butylsulfamoyl)-4-[2-[4- (isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate. 1H NMR (400MHz, METHANOL-d4) δ = 8.37 (d, J=2.2 Hz, 1H), 7.78 (s, 1H), 7.68 (br d, J=7.9 Hz, 1H), 7.40 (d, J=8.4 Hz, 1H), 4.86 - 4.78 (m, 1H), 4.11 (d, J=6.2 Hz, 2H), 3.50 - 3.39 (m, 3H), 3.11 - 2.96 (m, 3H), 2.23 (br d, J=12.3 Hz, 2H), 2.13 - 1.98 (m, 5H), 1.70 (dq, J=2.8, 12.8 Hz, 2H), 1.60 - 1.34 (m, 4H), 1.22 (br d, J=6.2 Hz, 6H), 1.11 (s, 9H). ESI [M/2+H] = 318.6
Example 34. Synthesis of isopropyl ((lR,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((R)- l-methylpyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2- yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000089_0003
Ex. 34
1H NMR (400MHz, METHANOL-d4) δ = 8.41 (d, J=2.2 Hz, 1H), 7.80 - 7.73 (m, 2H), 7.45 (d, J=8.4 Hz, 1H), 4.86 - 4.81 (m, 1H), 4.67 (dd, J=3.2, 12.8 Hz, 1H), 4.35 (dd, J=7.1, 12.8 Hz, IH), 3.85 - 3.71 (m, 2H), 3.47 (tt, J=3.9, 11.6 Hz, IH), 3.26 (td, J=8.1, 11.5 Hz, IH), 3.10 (s, 3H), 3.08 - 2.98 (m, IH), 2.46 - 2.35 (m, IH), 2.30 - 2.18 (m, 3H), 2.15 - 1.95 (m, 4H), 1.72 (dq, J=3.1, 12.9 Hz, 2H), 1.43 (dq, J=3.3, 12.6 Hz, 2H), 1.25 (br d, J=6.2 Hz, 6H), 1.13 (s, 9H). ESI [M+H] = 636.3
Example 35. Synthesis of isopropyl ((lS,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((S)-l- methylpyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2- yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000090_0001
Ex. 36
1H NMR (400MHz, METHANOL-d4) δ = 8.41 (d, J=2.0 Hz, IH), 7.80 - 7.71 (m, 2H), 7.44 (d, J=8.3 Hz, IH), 4.86 - 4.79 (m, IH), 4.67 (dd, J=3.2, 13.0 Hz, IH), 4.40 - 4.31 (m, IH), 3.87 - 3.66 (m, 2H), 3.47 (tt, J=3.8, 11.6 Hz, IH), 3.26 (td, J=8.3, 11.4 Hz, IH), 3.10 (s, 3H), 3.04 (tt, J=3.4, 12.0 Hz, IH), 2.48 - 2.33 (m, IH), 2.30 - 2.16 (m, 3H), 2.15 - 1.92 (m, 4H), 1.72 (dq, J=2.7, 12.8 Hz, 2H), 1.51 - 1.36 (m, 2H), 1.25 (br d, J=6.4 Hz, 6H), 1.13 (s, 9H). ESI [M/2+H] = 318.6
Example 36. Synthesis of isopropyl ((lR,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((R)- pyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000090_0002
Ex. 36
1H NMR (400MHz, METHANOL-d4) δ = 8.38 (br d, J=2.2 Hz, IH), 7.75 - 7.66 (m, 2H), 7.41 (d, J=8.4 Hz, IH), 4.81 (br s, IH), 4.48 (br dd, J=3.4, 12.5 Hz, IH), 4.33 (dd, J=7.8, 12.5 Hz, IH), 3.93 (dq, J=3.6, 8.0 Hz, IH), 3.49 - 3.33 (m, 3H), 3.00 (ddd, J=3.5, 8.5, 12.0 Hz, IH), 2.30 - 2.18 (m, 3H), 2.10 - 2.00 (m, 3H), 1.85 (qd, J=8.5, 12.9 Hz, 2H), 1.74 - 1.61 (m, 2H), 1.46 - 1.34 (m, 2H), 1.22 (br d, J=6.2 Hz, 6H), 1.09 (s, 9H). ESI [M/2+H] = 311.6
Example 37. Synthesis of isopropyl ((lS,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((S)- pyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000091_0001
Ex. 37
1H NMR (400MHz, METHANOL-d4) δ = 8.38 (d, J=1.8 Hz, IH), 7.79 - 7.68 (m, 2H), 7.41 (d, J=8.2 Hz, IH), 4.85 - 4.78 (m, IH), 4.49 (dd, J=3.4, 12.5 Hz, IH), 4.33 (dd, J=7.9, 12.3 Hz, IH), 3.94 (dq, J=3.4, 8.0 Hz, IH), 3.49 - 3.40 (m, IH), 3.40 - 3.33 (m, 2H), 3.08 - 2.97 (m, IH), 2.31 - 2.19 (m, 3H), 2.17 - 2.00 (m, 4H), 1.85 (qd, J=8.5, 13.0 Hz, IH), 1.75 - 1.62 (m, 2H), 1.46 - 1.35 (m, 2H), 1.22 (br d, J=6.2 Hz, 6H), 1.10 (s, 9H). ESI [M/2+H] = 311.6
Example 38. Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-
(((oxetan-3-yloxy)carbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000091_0002
Ex. 38
1H NMR (METHANOL-d4, 400MHz): δ = 8.33 (s, IH), 7.63-7.72 (m, 2H), 7.38 (d, J=8.4 Hz, IH), 5.50 (br t, J=5.5 Hz, IH), 4.92 (t, J=6.9 Hz, 2H), 4.81-4.83 (m, IH), 4.64-4.73 (m, 2H), 3.44 (br t, J=11.7 Hz, IH), 2.99 (br t, J=11.8 Hz, IH), 2.21 (br d, J=12.3 Hz, 2H), 2.06 (br d, J=11.7 Hz, 2H), 1.59-1.75 (m, 2H), 1.30-1.47 (m, 2H), 1.21 (br d, J=6.0 Hz, 6H), 1.10 ppm (s, 9H). ESI [M+H] = 595.1 Example 39. Synthesis of benzyl (3-(N-(tert-butyl)sulfamoyl)-4-(2-((lr,4r)-4-
((isopropoxycarbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000092_0001
Ex. 39
1H NMR (400MHz, METHANOL-d4) δ = 8.36 (s, IH), 7.75 - 7.67 (m, 2H), 7.46 - 7.28 (m, 6H), 5.28 - 5.14 (m, 2H), 4.85 - 4.76 (m, IH), 3.47 (br d, J=11.8 Hz, IH), 3.01 (br s, IH), 2.23 (br d, J=12.7 Hz, 2H), 2.07 (br d, J=14.5 Hz, 2H), 1.70 (br d, J=11.0 Hz, 2H), 1.40 (br d, J=12.7 Hz, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.12 (s, 9H). ESI [M+H] = 629.2
Example 40. Synthesis of 2-fluorobenzyl (3-(N-(tert-butyl)sulfamoyl)-4-(2-((lr,4r)-4-
((isopropoxycarbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000092_0002
Ex. 40
IH NMR (400MHz, METHANOL-d4) δ = 8.36 (s, IH), 7.77 - 7.63 (m, 2H), 7.52 (t, J=7.0 Hz, IH), 7.44 - 7.31 (m, 2H), 7.23 - 7.08 (m, 2H), 5.29 (s, 2H), 4.83 (br s, IH), 3.46 (br d, J=11.8 Hz, IH), 3.00 (br t, J=11.8 Hz, IH), 2.22 (br d, J=12.7 Hz, 2H), 2.07 (br d, J=11.4 Hz, 2H), 1.76 - 1.62 (m, 2H), 1.47 - 1.35 (m, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.12 (s, 9H). ESI [M+H] = 647.2 Example 41. Synthesis of (S)-l-phenylethyl (3-(N-(tert-butyl)sulfamoyl)-4-(2-((lr,4S)-4-
((isopropoxycarbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000093_0001
Ex. 41
1H NMR (400MHz, METHANOL-d4) δ = 8.35 (br s, IH), 7.81 - 7.65 (m, 2H), 7.48 - 7.36 (m, 5H), 7.31 (br d, J=6.8 Hz, IH), 5.89 (br d, J=6.2 Hz, IH), 4.85 (br d, J=5.5 Hz, IH), 3.47 (br s, IH), 3.03 (br s, IH), 2.24 (br d, J=11.2 Hz, 2H), 2.08 (br d, J=11.0 Hz, 2H), 1.71 (q, J=11.9 Hz, 2H), 1.61 (br d, J=6.4 Hz, 3H), 1.49 - 1.37 (m, 2H), 1.24 (br d, J=5.4 Hz, 6H), 1.13 (s, 9H). ESI [M+H] =643.2
Example 42. Synthesis of pyridin-2-ylmethyl (3-(N-(tert-butyl)sulfamoyl)-4-(2-((lr,4r)-
4-((isopropoxycarbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000093_0002
Ex. 42
1H NMR (400MHz, METHANOL-d4) δ = 8.56 (br d, J=4.5 Hz, IH), 8.38 (d, J=1.8 Hz, IH), 7.94 - 7.87 (m, IH), 7.78 - 7.72 (m, 2H), 7.59 (br d, J=7.8 Hz, IH), 7.44 - 7.38 (m, 2H), 5.32 (s, 2H), 4.85 (td, J=5.9, 12.0 Hz, IH), 3.47 (br t, J=11.8 Hz, IH), 3.07 - 2.96 (m, IH), 2.24 (br d, J=12.3 Hz, 2H), 2.13 - 2.04 (m, 2H), 1.77 - 1.65 (m, 2H), 1.48 - 1.37 (m, 2H), 1.24 (br d, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H] =630.2 Example 43. Synthesis of (R)-l-phenylethyl (3-(N-(tert-butyl)sulfamoyl)-4-(2-((lr,4R)-4-
((isopropoxycarbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate.
The following compound was synthesized via same method by the key intermediate 118.
Figure imgf000094_0001
1H NMR (400MHz, DMSO-d6) δ = 8.28 (d, J=2.2 Hz, IH), 7.70 - 7.57 (m, 2H), 7.47 - 7.24 (m, 7H), 7.05 - 6.84 (m, 2H), 5.82 (q, J=6.6 Hz, IH), 4.79 - 4.65 (m, IH), 3.35 - 3.24 (m, IH), 2.88 (tt, J=3.4, 11.9 Hz, IH), 2.11 (br d, J=11.7 Hz, 2H), 1.89 (br d, J=9.9 Hz, 2H), 1.54 (d, J=6.6 Hz, 5H), 1.30 (br s, 2H), 1.14 (d, J=6.2 Hz, 6H), 1.08 - 0.99 (m, 9H). ESI [M+H] = 643.3
Example 44. Synthesis of trans-isopropyl N-[4-[2-[4-(tert- butoxycarbonylamino )cyclohexyl] thiazol-5-yl]-3-( tert-butylsulfamoyl)pheny I] carbamate. Scheme 14:
Figure imgf000094_0002
82 Ex. 44
General method B, trans-isopropyl N-[4-[2-[4-(tert-butoxycarbonylamino)cyclohexyl] thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate. 1H NMR (400MHz, DMSO-d6) δ 1 0. Oil (s, IH), 8.32 (d, J=2.1 Hz, IH), 7.70 - 7.60 (m, 2H), 7.38 (d, J=8.4 Hz, IH), 6.96 (s, IH), 6.81 (br d, J=7.9 Hz, IH), 4.99 - 4.87 (m, IH), 3.27 (br s, IH), 2.94 - 2.84 (m, IH), 2.13 (br d, J=11.7 Hz, 2H), 1.90 (br d, J=l l. l Hz, 2H), 1.63 - 1.49 (m, 2H), 1.39 (s, 9H), 1.33 (br d, J=14.4 Hz, 2H), 1.28 (d, J=6.4 Hz, 6H), 1.07 (s, 9H). ESI [M+H] =595.1 Example 45. Synthesis of tert-butyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3- (pyridin-2-ylmethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 82.
Figure imgf000095_0001
Ex. 45
1H NMR (400MHz, METHANOL-d4) δ = 8.17 (d, J=2.2 Hz, 1H), 7.75 - 7.59 (m, 2H), 7.43 - 7.22 (m, 3H), 7.21 - 7.03 (m, 2H), 4.53 (m, 2H), 3.43 (br t, J=11.8 Hz, 1H), 2.99 (br t, J=12.1 Hz, 1H), 2.22 (br d, J=12.7 Hz, 2H), 2.06 (br d, J=10.5 Hz, 2H), 1.77 - 1.60 (m, 2H), 1.58 - 1.34 (m, 11H), 1.09 (s, 9H). ESI [M+H] =643.3
Example 46. Synthesis of isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[3-(isopropoxycarbonyl amino)azetidin-l-yl]thiazol-5-yl]phenyl]carbamate.
Scheme 15:
HN, NHBoc
Figure imgf000095_0002
128 129
Figure imgf000095_0003
130 Ex. 46
Preparation of compound 129.
tert-amyl alcohol, 100°C
Figure imgf000095_0004
128 129 A mixture of tert-butyl N-(azetidin-3-yl)carbamate;hydrochloride (72.40 mg, 346.94 umol, 1.50 eq.) , N-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl)phenyl] acetamide (100 mg, 231.29 umol, 1 eq.), t-BuONa (66.68 mg, 693.87 umol, 3 eq.) , [2-(2-aminoethyl)phenyl]- chloro-palladium;ditert-butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (15.88 mg, 23.13 umol, 0.1 eq.) in tert-amyl alcohol (2 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 100°C for 12 hrs under N2 atmosphere and then concentrated. The residue was diluted with Ethyl acetate (20 mL) and washed with H20 (20 mL). The organic layer was dried and concentrated and the residue was purified by prep- TLC (EtOAc) to give tert-butyl N-[l-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl]thiazol- 2-yl] azetidin-3-yl]carbamate (46 mg, crude) as a yellow solid. ESI [M+H] =524.3
Preparation of compound 130.
Figure imgf000096_0001
129 130
General method F, 5-amino-2-[2-(3-aminoazetidin-l-yl)thiazol-5-yl]-N-tert-butyl■ benzenesulfonamide. ESI [M+H] =382.0
Preparation of Example 46
Figure imgf000096_0002
130 Ex. 46
General method D, isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[3-(isopropoxycarbonyl amino)azetidin-l-yl]thiazol-5-yl]phenyl]carbamate. 1H NMR (400MHz, METHANOL-d4) δ = 8.35 (d, J=2.0 Hz, 1H), 7.67 (dd, J=2.2, 8.4 Hz, 1H), 7.45 - 7.32 (m, 2H), 5.04 - 4.94 (m, 1H), 4.92 - 4.88 (m, 1H), 4.73 - 4.63 (m, 1H), 4.56 (br t, J=8.5 Hz, 2H), 4.27 (br s, 2H), 1.31 (d, J=6.2 Hz, 6H), 1.27 - 1.19 (m, 15H). ESI [M+H] =554.2 Example 47. Synthesis οΐ trans-oxetan-3-yl N-[4-[5-[4-(benzylcarbamoylamino)-2-(tert- butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate
Scheme 16:
Figure imgf000097_0001
Ex. 47
Preparation of compound 131.
Figure imgf000097_0002
82 131
General method F, trans-4-(5-bromothiazol-2-yl)cyclohexanamine. ESI [M+H]
=260.9/262.9
Preparation of compound 132.
Figure imgf000097_0003
131 132
To a solution of oxetan-3-ol (1.15 g, 15.51 mmol, 3 eq.) in DCE (10 mL) were added DIEA (3.34 g, 25.84 mmol, 4.50 mL, 5 eq.) and TRIPHOSGENE (1.53 g, 5.17 mmol, 1 eq.). The mixture was stirred at 25-50°C for 1 hr and then added a solution of trans-4-(5-bromothiazol- 2-yl)cyclohexanamine (1.35 g, 5.17 mmol, 1 eq.), DIEA (3.34 g, 25.84 mmol, 4.50 mL, 5 eq.) in DCE (10 mL). The mixture was stirred at 25°C for 0.5 hr. The mixture was concentrated and the residue was purified by column chromatography (Si02, Petroleum ether/Ethyl acetate=30/l to 5: 1) to give trans-oxetan-3-yl N-[4-(5-bromothiazol-2- yl)cyclohexyl]carbamate (1.5 g, crude) as a white solid. ESI [M+H] =363.1/361.1
Figure imgf000098_0001
General method K, trans-oxetan-3-yl N-[4-[5-[4-(benzylcarbamoylamino)-2-(tert- butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate. 1H NMR (400MHz,
METHANOL-d4) δ = 8.23 (d, J=2.2 Hz, IH), 7.71 - 7.66 (m, 2H), 7.37 - 7.29 (m, 5H), 7.24 (br s, IH), 5.40 - 5.27 (m, 2H), 4.66 - 4.55 (m, 3H), 4.40 (s, 2H), 3.45 (br d, J=11.0 Hz, IH), 3.00 (br t, J=11.8 Hz, IH), 2.22 (br d, J=13.0 Hz, 2H), 2.07 (br d, J=11.2 Hz, 2H), 1.74 - 1.63 (m, 2H), 1.47 - 1.37 (m, 2H), 1.10 (s, 9H). ESI [M+H] =642.3
Example 48. Synthesis of oxetan-3-yl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-
((isopropoxycarbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 132.
Figure imgf000098_0002
Ex. 48
1H NMR (400MHz, METHANOL-d4) δ = 8.33 (d, J=2.0 Hz, IH), 7.71 - 7.64 (m, 2H), 7.36 (d, J=8.4 Hz, IH), 5.41 - 5.27 (m, 2H), 4.98 (td, J=6.4, 12.6 Hz, IH), 4.87 (br s, IH), 4.63 - 4.56 (m, 2H), 3.45 (br d, J=12.3 Hz, IH), 3.00 (br t, J=12.0 Hz, IH), 2.22 (br d, J=13.0 Hz, 2H), 2.11 - 2.02 (m, 2H), 1.74 - 1.63 (m, 2H), 1.42 (q, J=12.7 Hz, 2H), 1.31 (d, J=6.2 Hz, 6H), 1.11 (s, 9H). ESI [M+H] =595.3
Example 49. Synthesis oitrans-oxetan-3-yl N-[3-(tert-butylsulfamoyl)-4-[2-[4-(oxetan-3- yloxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate
Scheme 17:
Figure imgf000099_0001
Preparation of compound 133.
Figure imgf000099_0002
General method K, trans-oxetan-3-yl N-[4-[5-[4-amino-2-(tert-butylsulfamoyl) phenyl]thiazol-2-yl]cyclohexyl]carbamate. ESI [M+H] =509.0
Preparation of compound 134.
Figure imgf000099_0003
General method D, trans-oxetan-3-yl N-[4-[5-[2-(tert-butylsulfamoyl)-4- [(4- nitrophenoxy)carbonylamino]phenyl]thiazol-2-yl]cyclohexyl]carbamate. ESI [M+H] =674.2 Preparation of Example 49.
Figure imgf000100_0001
134 Ex. 49
General method H, trans-oxetan-3-yl N-[3-(tert-butylsulfamoyl)-4-[2-[4-(oxetan-3- yloxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate. 1H NMR (400MHz, DMSO-d6) δ = 10.39 (s, IH), 8.28 (d, J=2.2 Hz, IH), 7.71 - 7.58 (m, 2H), 7.48 - 7.34 (m, 2H), 6.97 (s, IH), 5.50 - 5.39 (m, IH), 5.32 - 5.20 (m, IH), 4.87 - 4.68 (m, 4H), 4.61 - 4.36 (m, 4H), 3.30 - 3.23 (m, IH), 3.01 - 2.82 (m, IH), 2.12 (br d, J=11.7 Hz, 2H), 1.90 (br d, J=10.4 Hz, 2H), 1.66 - 1.47 (m, 2H), 1.43 - 1.26 (m, 2H), 1.04 (s, 9H). ESI [M+H] =609.2
Example 50. Synthesis of oxetan-3-yl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3- (pyridin-2-ylmethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 134.
Figure imgf000100_0002
Ex. 50
1H NMR (400MHz, METHANOL-d4) δ = 8.49 (d, J=4.8 Hz, IH), 8.25 (d, J=2.2 Hz, IH), 7.82 (dt, J=1.8, 7.7 Hz, IH), 7.73 - 7.63 (m, 2H), 7.45 (d, J=7.9 Hz, IH), 7.38 - 7.26 (m, 2H), 5.39 - 5.28 (m, IH), 4.84 (br s, IH), 4.70 - 4.56 (m, 3H), 4.53 (s, 2H), 3.44 (br t, J=12.1 Hz, IH), 3.06 - 2.92 (m, IH), 2.22 (br d, J=13.2 Hz, 2H), 2.07 (br d, J=11.0 Hz, 2H), 1.79 - 1.60 (m, 2H), 1.50 - 1.35 (m, 2H), 1.10 (s, 9H). ESI [M+H] =643.2
Example 51. Synthesis of oxetan-3-yl ((lS,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((S)- l-phenylethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 134.
Figure imgf000101_0001
Ex. 51
1H NMR (400MHz, METHANOL-d4) δ = 8.19 (d, J=2.2 Hz, 1H), 7.70 - 7.63 (m, 2H), 7.39 - 7.30 (m, 5H), 7.27 - 7.20 (m, 1H), 5.35 (t, J=5.7 Hz, 1H), 4.92 (q, J=6.8 Hz, 2H), 4.84 (br s, 1H), 4.60 (t, J=6.4 Hz, 2H), 3.49 - 3.38 (m, 1H), 3.05 - 2.95 (m, 1H), 2.22 (br d, J=12.3 Hz, 2H), 2.10 - 2.02 (m, 2H), 1.74 - 1.61 (m, 2H), 1.49 (d, J=7.1 Hz, 3H), 1.45 - 1.34 (m, 2H), 1.09 (s, 9H). ESI [M+H] =656.3
Example 52. Synthesis of oxetan-3-yl ((lR,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3- ((R)-l-phenylethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 134.
Figure imgf000101_0002
1HNMR (400MHz, METHANOL-d4) δ = 8.19 (d, J=2.2 Hz, 1H), 7.75 - 7.61 (m, 2H), 7.42 - 7.18 (m, 6H), 5.35 (quin, J=5.7 Hz, 1H), 5.01 - 4.87 (m, 3H), 4.70 - 4.52 (m, 2H), 3.51 - 3.38 (m, 1H), 3.06 - 2.92 (m, 1H), 2.22 (br d, J=11.9 Hz, 2H), 2.07 (br d, J=11.7 Hz, 2H), 1.77 - 1.62 (m, 2H), 1.54 - 1.35 (m, 5H), 1.10 (s, 9H). ESI [M+H] =656.2 Example 53. Synthesis of oxetan-3-yl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(2- fluorobenzyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 134.
Figure imgf000102_0001
Ex. 53
1HNMR (400MHz, METHANOL-d4) δ = 8.23 (d, J=2.4 Hz, 1H), 7.75 - 7.63 (m, 2H), 7.46 - 7.24 (m, 3H), 7.20 - 7.03 (m, 2H), 5.43 - 5.28 (m, 1H), 4.85 (br s, 2H), 4.67 - 4.56 (m, 2H), 4.47 (s, 2H), 3.53 - 3.37 (m, 1H), 3.08 - 2.93 (m, 1H), 2.29 - 1.99 (m, 4H), 1.78 - 1.61 (m, 2H), 1.53 - 1.35 (m, 2H), 1.11 (s, 9H). ESI [M+H] =660.2
Example 54. Synthesis of oxetan-3-yl ((lR,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3- ((R)-l-(2-fluorophenyl)ethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 134.
Figure imgf000102_0002
1H NMR (400MHz, METHANOL-d4) δ = 8.20 (d, J=2.3 Hz, 1H), 7.73 - 7.65 (m, 2H), 7.45 - 7.39 (m, 1H), 7.38 - 7.26 (m, 2H), 7.21 - 7.04 (m, 2H), 5.37 (quin, J=5.7 Hz, 1H), 5.20 (q, J=7.0 Hz, 1H), 4.89 - 4.86 (m, 2H), 4.65 - 4.59 (m, 2H), 3.52 - 3.41 (m, 1H), 3.08 - 2.96 (m, 1H), 2.24 (br d, J=12.2 Hz, 2H), 2.13 - 2.05 (m, 2H), 1.76 - 1.64 (m, 2H), 1.52 (d, J=7.0 Hz, 3H), 1.47 - 1.39 (m, 2H), 1.12 (s, 9H). ESI [M+H] =674.2 Example 55. Synthesis of oxetan-3-yl ((lS,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((S)- l-(2-fluorophenyl)ethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate.
The following compound was synthesized via same method by the key intermediate 134.
Figure imgf000103_0001
Ex. 55
1HNMR (400MHz, METHANOL-d4) δ = 8.17 (d, J=2.2 Hz, 1H), 7.75 - 7.59 (m, 2H), 7.43 - 7.22 (m, 3H), 7.21 - 7.03 (m, 2H), 5.39 - 5.28 (m, 1H), 5.17 (q, J=7.0 Hz, 1H), 4.90 - 4.87 (m, 1H), 4.84 (s, 1H), 4.66 - 4.54 (m, 2H), 3.43 (br t, J=11.8 Hz, 1H), 2.99 (br t, J=12.1 Hz, 1H), 2.22 (br d, J=12.7 Hz, 2H), 2.06 (br d, J=10.5 Hz, 2H), 1.77 - 1.60 (m, 2H), 1.58 - 1.34 (m, 5H), 1.09 (s, 9H). ESI [M+H] =674.2
Example 56. Synthesis of 4-piperidylmethyl N-[3-(tert-butylsulfamoyl)-4-[2-[5- (isopropoxycarbonylamino)-3-methoxy-2-pyridyl]thiazol-5-yl]phenyl]carbamate
Scheme 18:
NH,
Figure imgf000103_0002
135 136 137 138
Figure imgf000103_0003
141 142
Figure imgf000104_0001
Figure imgf000104_0002
Ex. 56
Preparation of compound 136.
Figure imgf000104_0003
135 136
Na (8.75 g, 380.60 mmol, 9.02 mL, 1.51 eq.) was added into MeOH (300 mL) portionwise and after Na was dissolved, the mixture was concentrated to dryness. The resulting gray solid (NaOMe) was added into DMF (300 mL) and 3-chloropyridine-2-carbonitrile (35 g, 252.61 mmol, 1 eq.) was added at 0°C. The reaction mixture was stirred at 20°C for 12 hrs and then diluted with H20 (800 mL) and filtered. The cake dried to give 3-methoxypyridine- 2-carbonitrile (25 g, crude) as a white solid. ESI [M+H] =135.1
Preparation of compound 137
Figure imgf000104_0004
136 137
To a solution of 3-methoxypyridine-2-carbonitrile (24.5 g, 182.65 mmol, 1 eq.) in DCM (450 mL) was added a mixture of Bu4NN03 (83.30 g, 273.59 mmol, 1.5 eq.) and TFAA (57.54 g, 273.98 mmol, 38.11 mL, 1.5 eq.) in DCM (150 mL) dropwise at 0°C. The mixture was stirred at 20°C for 12 hrs and then poured into sat.aq.NaHC03 (300 mL) at 0°C and the organic layer was dried over Na2S04, filtered and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether/Ethyl acetate=20/l to 10: 1) to give 3- methoxy-5-nitro-pyridine-2-carbonitrile (27 g, crude) as a white solid. 1H NMR (400MHz, CHLOROFORM-d) δ = 9.06 (d, J=2.2 Hz, 1H), 8.13 (d, J=2.2 Hz, 1H), 4.16 - 4.08 (m, 3H). ESI [M+H] =180.1
General method G for preparation of compound 138.
Figure imgf000105_0001
137 138
To a solution of 3-methoxy-5-nitro-pyridine-2-carbonitrile (27 g, 151 mmol, 1 eq.) in THF (100 mL)/EtOH (500 mL) were added Fe (4 g, 754 mmol, 5 eq.) and a solution of NH4C1 (24.2 g, 452 mmol, 3 eq.) in H20 (50 mL). The mixture was stirred at 80°C for 30 mins, then diluted with THF (500 mL) and filtered. The filtrate was concentrated, diluted with H20 (500 mL) and then extracted with DCM (400 mL x 3). The combined organic layers were dried over Na2S04, filtered and concentrated to give 5-amino-3-methoxy-pyridine-2- carbonitrile (14.5 g, crude) as a pale yellow solid. ESI [M+H] =150.1
Preparation of compound 139.
Figure imgf000105_0002
138 139
General method D, isopropyl N-(6-cyano-5-methoxy-3-pyridyl)carbamate. 1H NMR (400MHz, METHANOL-d4) δ = 8.13 (d, J=2.2 Hz, 1H), 7.95 (d, J=1.8 Hz, 1H), 5.04 - 4.95 (m, 1H), 3.96 (s, 3H), 1.31 (d, J=6.2 Hz, 6H). ESI [M+H] =236.1 Preparation of compound 140.
Figure imgf000106_0001
To a solution of isopropyl N-(6-cyano-5-methoxy-3-pyridyl)carbamate (16 g, 68.02 mmol, 1 eq.) in DMF (200 mL) were added NaHS (19.06 g, 340.08 mmol, 5 eq.) and then MgCl2 (19.43 g, 204.05 mmol, 8.37 mL, 3 eq.). The mixture was stirred at 25°C for 12 hrs and then poured into H20 (500ml) and extracted with DCM (40 mL * 3). The combined organic layers were dried over Na2S04, filtered and concentrated. The residue was washed with a solution (Petroleum ether : EtOAc= 8: 1), filtered and the filter cake was dried to give isopropyl N-(6- carbam o th ioy I- 5- m eth oxy- 3-pyridyl)carbamate (21 g, crude) as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ = 9.92 - 9.84 (m, 2H), 9.38 (br s, 1H), 8.08 (d, J=1.8 Hz, 1H), 7.66 (s, 1H), 4.89 (spt, J=6.3 Hz, 1H), 3.74 (s, 3H), 1.24 (d, J=6.1 Hz, 6H). ESI [M+H] =270.0
Preparation of compound 141.
Figure imgf000106_0002
General method M, isopropyl N-(5-methoxy-6-thiazol-2-yl-3-pyridyl)carbamate 1H NMR (400MHz, DMSO-d6) δ = 10.07 (s, 1H), 8.26 (d, J=2.0 Hz, 1H), 7.88 (d, J=3.1 Hz, 1H), 7.85 (d, J=1.3 Hz, 1H), 7.71 (d, J=3.3 Hz, 1H), 4.91 (spt, J=6.2 Hz, 1H), 3.88 (s, 3H), 1.26 (d, J=6.4 Hz, 6H). ESI [M+H] =294.1
Preparation of compound 142.
Figure imgf000106_0003
General method J, isopropyl N-[6-(5-bromothiazol-2-yl)-5-methoxy-3-pyridyl] carbamate.
1H NMR (400MHz, DMSO-d6) δ = 10.17 (s, 1H), 8.28 (d, J=1.8 Hz, 1H), 8.08 - 7.86 (m, 2H), 4.99 (br s, 1H), 3.93 (s, 3H), 1.29 (d, J=6.4 Hz, 6H). ESI [M+H] =371.8/373.8
Preparation of compound 143.
Figure imgf000107_0001
142 143
General method B, isopropyl N-[6-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl] thiazol-2- yl]-5-methoxy-3-pyridyl]carbamate. ESI [M+H] =562.0
Preparation of compound 144.
Figure imgf000107_0002
143 144
General method F, isopropyl N-[6-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl] thiazol-2-yl]- 5-methoxy-3-pyridyl]carbamate. ESI [M+H] =520.2
Preparation of compound 145.
Figure imgf000107_0003
144 145
General method D, (4-nitrophenyl) N-[3-(tert-butylsulfamoyl)-4-[2-[5- (isopropoxycarbonylamino)-3-methoxy-2-pyridyl]thiazol-5-yl]phenyl]carbamate. ESI
[M+H] =685.1 Preparation of compound 146.
Figure imgf000108_0001
145 146
General method H, tert-butyl 4-[[3-(tert-butylsulfamoyl)-4-[2-[5- ( isopropoxycarbonylamino )-3-methoxy-2-pyridyl]thiazol-5- yl]phenyl]carbamoyloxymethyl]piperidine-l-carboxylate. ESI [M+H]
Preparation of Ex. 56
Figure imgf000108_0002
General method C, 4-piperidylmethyl N-[3-(tert-butylsulfamoyl)-4-[2-[5- (isopropoxycarbonylamino)-3-methoxy-2-pyridyl]thiazol-5-yl]phenyl]carbamate. 1H NMR
(400MHz, DMSO-d6) δ = 10.19 - 10.10 (m, 2H), 8.35 (d, J=2.1 Hz, IH), 8.30 (d, J=1.8 Hz, IH), 7.89 (s, 2H), 7.68 (dd, J=2.0, 8.4 Hz, IH), 7.46 (d, J=8.4 Hz, IH), 7.12 (br s, IH), 4.95 (td, J=6.2, 12.5 Hz, IH), 3.98 (d, J=6.5 Hz, 2H), 3.93 (s, 3H), 3.00 (br d, J=12.1 Hz, 2H), 2.70 - 2.56 (m, 2H), 1.77 (br d, J=4.0 Hz, IH), 1.67 (br d, J=12.1 Hz, 2H), 1.30 (d, J=6.2 Hz, 6H), 1.16 (br dd, J=3.4, 12.0 Hz, 2H), 1.09 (s, 9H). ESI [M+H] =661.3
Example 57 Synthesis of trans-isopropyl N-[4-[5-[4-(benzylcarbamoylamino)-2-(tert- butylsulfamoyl)phenyl]-4-fluoro-thiazol-2-yl]cyclohexyl]carbamate.
Scheme 19:
Figure imgf000108_0003
116 147
Figure imgf000109_0001
Preparation of compound 147
Figure imgf000109_0002
To a solution of trans-isopropyl N-[4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (1.50 g, 4.32 mmol, 1 eq.) in ACN (20 mL) was added l-(chloromethyl)-4-fluoro- l,4 - diazoniabicyclo[2.2.2]octane;ditetrafluoroborate (3.06 g, 8.64 mmol, 2 eq.) and the mixture was stirred at 80°C for 12 hrs. The mixture was then concentrated and the residue was purified by column chromatography (Si02, Petroleum ether/Ethyl acetate=3/l to 3: 1) to give trans-isopropyl N-[4-(5-bromo-4-fluoro-thiazol-2-yl) cyclohexyljcarbamate (0.2 g, 547.55 umol, 12.68% yield) as yellow gum. ESI [M+H] =365.1/367.1
Preparation of Ex. 57
Figure imgf000109_0003
General method B, trans-isopropyl N-[4-[5-[4-(benzylcarbamoylamino)-2-(tert- butylsulfamoyl)phenyl]-4-fluoro-thiazol-2-yl]cyclohexyl]carbamate. 1H NMR (400MHz, METHANOL-d4) δ = 8.24 (d, J=2.0 Hz, IH), 7.67 (dd, J=2.0, 8.4 Hz, IH), 7.37 - 7.29 (m, 5H), 7.24 (br d, J=2.6 Hz, IH), 4.82 - 4.77 (m, IH), 4.40 (s, 2H), 3.42 (br t, J=l 1.6 Hz, IH), 2.87 (br t, J=12.0 Hz, IH), 2.20 (br d, J=12.3 Hz, 2H), 2.05 (br d, J=11.0 Hz, 2H), 1.69 - 1.57 (m, 2H), 1.43 - 1.30 (m, 2H), 1.21 (br d, J=5.7 Hz, 6H), 1.15 (s, 9H). ESI [M+H] =646.2 Example 58 Synthesis of isopropyl ((lr,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4- ((isopropoxycarbonyl)amino)phenyl)-4-fluorothiazol-2-yl)cyclohexyl)carbamate
The following compound was synthesized via same method by the key intermediate 147.
Figure imgf000110_0001
1H NMR (400MHz, METHANOL-d4) δ = 8.35 (s, 1H), 7.69 (br d, J=8.1 Hz, 1H), 7.37 (d, J=8.3 Hz, 1H), 5.01 (td, J=6.1, 12.4 Hz, 1H), 4.83 (br s, 1H), 3.53 - 3.39 (m, 1H), 2.96 - 2.85 (m, 1H), 2.23 (br d, J=12.3 Hz, 2H), 2.08 (br d, J=10.5 Hz, 2H), 1.75 - 1.60 (m, 2H), 1.47 - 1.38 (m, 2H), 1.34 (d, J=6.2 Hz, 6H), 1.24 (br d, J=6.0 Hz, 6H), 1.18 (s, 9H). ESI [M+H] =599.2
Example 59 Synthesis of trans-isopropyl N-[ 6-[5-[4-(benzylcarbamoylamino ) -2-(tert- butylsulfamoyl)phenyl]thiazol-2-yl]tetrahydropyran-3-yl]carbamate
Scheme 20:
Figure imgf000110_0002
157 158
Figure imgf000111_0001
Figure imgf000111_0002
Preparation of compoundl49
Figure imgf000111_0003
148 149
To a solution of ethyl 2-oxoacetate (250 g, 1.22 mol, 1 eq.) in Tol. (1.5 L), were added buta- 1,3-diene (92.72 g, 1.71 mol, 149.55 mL, 1.4 eq.) and 2,6-ditert-butyl-4-methyl-phenol (5.40 g, 24.49 mmol, 0.02 eq.). The mixture was stirred at 170°C for 8 hrs in high pressure tube under 1 MPa and then concentrated. The residue was purified by silica gel chromatography (Petroleum ether/Ethyl acetate=100: 1- 10: 1) to afford ethyl 3,6-dihydro-2H-pyran-2- carboxylate (26 g, 166.48 mmol, 13.60% yield) as yellow oil. 1H NMR (400MHz,
CHLOROFORM-d) δ = 5.82 - 5.74 (m, 1H), 5.71 - 5.64 (m, 1H), 4.36 - 4.27 (m, 1H), 4.23 - 4.12 (m, 4H), 2.39 - 2.23 (m, 2H), 1.24 (t, J=7.1 Hz, 3H).
Preparation of compound 150
Figure imgf000111_0004
149 150
General method O, 3,6-dihydro-2H-pyran-2-carboxylic acid. 1H NMR (400MHz,
CHLOROFORM-d) δ = 5.88 (qdd, J=2.3, 5.1, 10.2 Hz, 1H), 5.80 - 5.71 (m, 1H), 4.43 - 4.19 (m, 3H), 2.53 - 2.31 (m, 2H) Preparation of compound 151
Figure imgf000112_0001
50 151
General method N, 3,6-dihydro-2H-pyran-2-carboxamide. 1H NMR (400MHz,
CHLOROFORM-d) δ = 6.50 (br s, IH), 5.89 - 5.76 (m, IH), 5.67 (br d, J=9.8 Hz, IH), 5.50 (br s, IH), 4.22 (br s, 2H), 3.97 (dd, J=3.9, 10.8 Hz, IH), 2.49 - 2.35 (m, IH), 2.28 - 2.09 (m, IH)
Preparation of compound 152
Figure imgf000112_0002
General method L, 3,6-dihydro-2H-pyran-2-carbothioamide. ESI [M+H]=144.1 Preparation of compound 153
Figure imgf000112_0003
152 153
General method M, 2-(3,6-dihydro-2H-pyran-2-yl)thiazole. 1H NMR (400MHz,
CHLOROFORM-d) δ = 7.69 (d, J=2.9 Hz, IH), 7.26 (d, J=2.9 Hz, IH), 5.93 - 5.83 (m, IH), 5.70 - 5.63 (m, IH), 4.86 (dd, J=3.9, 9.8 Hz, IH), 4.33 (br s, 2H), 2.61 - 2.37 (m, 2H). ESI [M+H]=168.1
Preparation of compound 154
Figure imgf000112_0004
153 154 To a solution of 2-(3,6-dihydro-2H-pyran-2-yl)thiazole (13 g, 77.74 mmol, 1 eq.) in THF (150 mL), was added BH3-Me2S (10 M, 15.55 mL, 2 eq.) at 0 °C dropwise and the mixture was stirred at 26°C for 2 hrs. Then the mixture was quenched by NaOH (62.19 g, 1.55 mol, 20 eq.) in H20 (150 mL) slowly at 0 °C followed by addition of H202 (264.42 g, 2.33 mol, 224.09 mL, 30% purity, 30 eq.). The mixture was stirred at 26°C for another 12 hrs. The mixture was quenched by sat.aq.Na2S03 solution (1 L) and extracted with EtOAc (1 L*2). The combined organic phased was dried over Na2S04, filtered and concentrated. The residue was purified by column (Petroleum ether : EtOAc= 5: 1- 1 : 1) to afford 6-thiazol-2- yltetrahydropyran-3-ol (7 g, crude) as a yellow solid. ESI [M+H]=186.2
Preparation of compound 155
Figure imgf000113_0001
154 155
To a solution of 6-thiazol-2-yltetrahydropyran-3-ol (7 g, 37.79 mmol, 1 eq.) in DCM (100 mL), were added TEA (7.65 g, 75.58 mmol, 10.52 mL, 2 eq.) and methanesulfonyl chloride (6.49 g, 56.68 mmol, 4.39 mL, 1.5 eq.) dropwise at 0 °C and the mixture was stirred at 26°C for 2 hrs. The mixture was diluted with DCM (100 mL) and washed with water (200 mL). The organic phase was dried over Na2S04, filtered and concentrated to give crude (6-thiazol- 2-yltetrahydropyran-3-yl) methanesulfonate (7 g, crude) as yellow oil which can be used directly.
To a solution of (6-thiazol-2-yltetrahydropyran-3-yl) methanesulfonate (7 g, 26.58 mmol, 1 eq.) in DMF (60 mL), was added azidosodium (8.64 g, 132.91 mmol, 5 eq.) and the mixture was stirred at 80°C for 12 hrs and then poured into sat.aq.Na2C03 (500 mL) and extracted with EtOAc (200 mL*3). The combined organic phase was washed with brine (200 mL) and dried over Na2S04, filtered and concentrated to give crude 2-(5-azidotetrahydropyran-2- yl)thiazole (5 g, crude) as yellow oil which can be used without any purification. Preparation of compound 156
Figure imgf000114_0001
155 156
To a solution of 2-(5-azidotetrahydropyran-2-yl)thiazole (5 g, 23.78 mmol, 1 eq.) in THF (80 mL) and H20 (40 mL), was added PPI13 (9.36 g, 35.67 mmol, 1.5 eq.). The mixture was stirred at 50°C for 12 hrs and then poured into 4N HCl solution (100 mL) and extracted with EtOAc (50 mL*2). Then the aqueous phase was bacified by sat.aq.Na2C03 until pH>12 and extracted with a solution (DCM/MeOH = 5: 1) (100 mL*3). The combined organic phase was dried over Na2S04, filtered and concentrated to give crude 6-thiazol-2-yltetrahydropyran-3- amine (3 g, crude) as yellow oil. ESI [M+H] =185.2
Preparation of compound 157
Figure imgf000114_0002
157
General method D, trans-isopropyl N-(6-thiazol-2-yltetrahydropyran-3-yl)carbamate. 1H
NMR (400MHz, METHANOL-d4) δ = 7.74 (d, J=3.3 Hz, 1H), 7.55 (d, J=3.3 Hz, 1H), 4.86 - 4.77 (m, 2H), 4.64 (dd, J=2.4, 10.8 Hz, 1H), 4.12 (ddd, J=2.0, 4.6, 10.8 Hz, 1H), 3.75 - 3.55 (m, 1H), 2.32 - 2.20 (m, 1H), 2.16 - 2.03 (m, 1H), 1.81 - 1.54 (m, 2H), 1.22 (br d, J=6.2 Hz, 6H). ESI [M+H] = 271.2
Note: Cpd.157 was purified by prep-TLC and then prep-HPLC to separate out other isomers.
Preparation of compound 158
Figure imgf000114_0003
H H
157 158 General method J, trans-isopropyl N-[6-(5-bromothiazol-2-yl)tetrahydropyran-3-yl] carbamate. ESI [M+H]=351.1/349.1
Preparation of compound Ex. 59
Figure imgf000115_0001
158
Ex. 59
General method B, trans-isopropyl N-[6-[5-[4-(benzylcarbamoylamino) -2-(tert- butylsulfamoyl)phenyl]thiazol-2-yl]tetrahydropyran-3-yl]carbamate. 1H NMR (400MHz, METHANOL-d4) δ = 8.24 (d, J=2.2 Hz, IH), 7.74 (s, IH), 7.68 (dd, J=2.2, 8.4 Hz, IH), 7.37 - 7.29 (m, 5H), 7.27 - 7.19 (m, IH), 4.84 - 4.78 (m, 2H), 4.64 (dd, J=2.0, 11.0 Hz, IH), 4.40 (s, 2H), 4.12 (br dd, J=3.2, 10.7 Hz, IH), 3.69 - 3.58 (m, IH), 2.33 - 2.26 (m, IH), 2.13 (br d, J=10.4 Hz, IH), 1.82 - 1.71 (m, IH), 1.69 - 1.57 (m, IH), 1.26 - 1.19 (m, 6H), 1.10 (s, 9H). ESI [M+H] =630.3
Preparation of compound Ex. 59 A and Ex. 59B
Figure imgf000115_0002
Ex. 59 was further separated by SFC (condition: Instrument: Thar SFC80 preparative SFC; Column: Chiralpak IC-H 250*30mm i.d. 5u; Mobile phase: A for C02 and B for
MeOH(0.1%NH3.H2O); Gradient: B%=42% ; Flow rate70g/min; Wavelength:220 nni; Column temperature: 40 °C ; System back pressure: 100 bar to give Ex. 59A. 1H NMR (400MHz, METHANOL-d4) δ = 8.27 (d, J=2.2 Hz, IH), 7.77 (s, IH), 7.72 (dd, J=2.3, 8.3 Hz, IH), 7.40 - 7.33 (m, 5H), 7.30 - 7.23 (m, IH), 4.89 - 4.79 (m, 2H), 4.67 (dd, J=2.3, 11.0 Hz, IH), 4.43 (s, 2H), 4.16 (br dd, J=3.0, 10.9 Hz, IH), 3.67 (br t, J=10.9 Hz, IH), 2.33 (br dd, J=2.6, 13.1 Hz, IH), 2.16 (br d, J=10.6 Hz, IH), 1.88 - 1.58 (m, 2H), 1.31 - 1.22 (m, 6H), 1.13 (s, 9H). ESI [M+H] =630.2
Ex. 59B: 1H NMR (400MHz, METHANOL-d4) δ = 8.25 (d, J=2.2 Hz, IH), 7.76 (s, IH), 7.70 (dd, J=2.2, 8.3 Hz, IH), 7.37 (s, IH), 7.36 - 7.31 (m, 4H), 7.28 - 7.18 (m, IH), 4.87 - 4.74 (m, IH), 4.65 (br d, J=9.2 Hz, IH), 4.41 (s, 2H), 4.19 - 4.04 (m, IH), 3.63 (br d, J=10.5 Hz, IH), 3.34 (br s, IH), 2.31 (br d, J=13.2 Hz, IH), 2.13 (br d, J=10.1 Hz, IH), 1.84 - 1.72 (m, IH), 1.71 - 1.58 (m, IH), 1.22 (d, J=6.1 Hz, 6H), 1.11 (s, 9H). ESI [M+H] =630.3
Example 60 Synthesis of trans-isopropyl N-[6-[5-[2-(tert-butylsulfamoyl)-4- (isopropoxycarbonylamino)phenyl]thiazol-2-yl]tetrahydropyran-3-yl]carbamate
Scheme 21:
Figure imgf000116_0001
Figure imgf000116_0002
Preparation of Ex. 60
Figure imgf000116_0003
General method B, trans-isopropyl N-[6-[5-[2-(tert-butylsulfamoyl)-4- (isopropoxycarbonylamino)phenyl]thiazol-2-yl]tetrahydropyran-3-yl]carbamate. 1H NMR
(400MHz, METHANOL-d4) δ = 8.37 (d, J=2.0 Hz, IH), 7.79 (s, IH), 7.70 (dd, J=2.4, 8.3 Hz, IH), 7.40 (d, J=8.3 Hz, IH), 5.01 (spt, J=6.3 Hz, IH), 4.87 - 4.79 (m, IH), 4.68 (dd, J=2.4, 11.2 Hz, IH), 4.16 (dd, J=2.9, 10.8 Hz, IH), 3.72 - 3.60 (m, IH), 3.38 - 3.34 (m, IH), 2.39 - 2.29 (m, IH), 2.16 (br d, J=12.2 Hz, IH), 1.89 - 1.74 (m, IH), 1.72 - 1.59 (m, IH), 1.39 - 1.21 (m, 12H), 1.14 (s, 9H). ESI [M+H] =583.3
Preparation of compound Ex. 60 and Ex. 60B
Figure imgf000117_0001
Ex. 60B
Ex. 60 was further separated by SFC (condition: Instrument: Thar SFC80 preparative SFC; Column: Chiralpak IC-H 250*30mm i.d. 5u; Mobile phase: A for C02 and B for
MeOH(0.1%NH3.H2O); Gradient: B%=38% ; Flow rate:65g/min; Wavelength:220 nni; Column temperature: 40 °C ; System back pressure: 100 bar) to give Ex. 60A 1H NMR (400MHz, METHANOL-d4) δ = 8.37 (d, J=2.2 Hz, IH), 7.78 (s, IH), 7.70 (dd, J=2.0, 8.4 Hz, IH), 7.40 (d, J=8.3 Hz, IH), 5.05 - 4.95 (m, IH), 4.90 - 4.81 (m, IH), 4.67 (dd, J=2.4, 11.1 Hz, IH), 4.70 - 4.64 (m, IH), 4.16 (br dd, J=2.9, 10.9 Hz, IH), 3.73 - 3.62 (m, IH), 2.33 (br dd, J=2.8, 13.2 Hz, IH), 2.16 (br d, J=11.9 Hz, IH), 1.86 - 1.74 (m, IH), 1.70 - 1.61 (m, IH), 1.34 (d, J=6.2 Hz, 6H), 1.25 (br d, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H] =583.3
Ex. 60B: 1H NMR (400MHz, METHANOL-d4) δ = 8.34 (d, J=2.2 Hz, IH), 7.80 - 7.72 (m, IH), 7.67 (dd, J=2.0, 8.6 Hz, IH), 7.36 (d, J=8.3 Hz, IH), 4.98 (spt, J=6.2 Hz, IH), 4.85 - 4.75 (m, IH), 4.65 (dd, J=2.4, 11.2 Hz, IH), 4.13 (br dd, J=3.1, 11.0 Hz, IH), 3.72 - 3.56 (m, IH), 3.33 (s, IH), 2.30 (br dd, J=2.6, 13.2 Hz, IH), 2.13 (br d, J=11.4 Hz, IH), 1.85 - 1.71 (m, IH), 1.69 - 1.57 (m, IH), 1.31 (d, J=6.1 Hz, 6H), 1.27 - 1.17 (m, 6H), 1.11 (s, 9H). ESI [M+H] =583.2 Example 61 Synthesis of [l-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl] thiazol-2-yl]-4-bicyclo[2.2.2]octanyl] N-isopropylcarbamate.
Scheme 22:
MeOOC
Figure imgf000118_0001
159 160
Figure imgf000118_0002
162 163
Figure imgf000118_0003
164 165
Figure imgf000118_0004
Ex. 61
Preparation of compound 160.
Figure imgf000118_0005
159 160
To a solution of methyl 4-hydroxybicyclo[2.2.2]octane-l-carboxylate (0.4 g, 2.17 mmol, 1 eq.) in DCM (10 mL) were added TMSCI (23.59 mg, 217.12 umol, 0.1 eq.) and 2- isocyanatopropane (554.33 mg, 6.51 mmol, 3 eq.). The mixture was stirred at 25°C for 12 hrs and then washed with 1 N HC1 (20 mL) and sat.aq.Na2C03 (20 mL). The organic layer was dried over Na2S04, filtered and concentrated to give methyl 4- (isopropylcarbamoyloxy)bicyclo[2.2.2]octane-l- carboxylate (0.45 g, crude) as a yellow gum. 1H NMR (400MHz, CHLOROFORM-d) δ = 4.34 (br s, 1H), 3.72 - 3.61 (m, 1H), 3.56 (s, 3H), 1.94 (br d, J=7.7 Hz, 6H), 1.90 - 1.81 (m, 6H), 1.08 - 1.00 (m, 6H). ESI [M+H] =269.9
Preparation of compound 161.
Figure imgf000119_0001
160 161
Gerneral method O, 4-(isopropylcarbamoyloxy)bicyclo[2.2.2]octane-l-carboxylic acid. 1H
NMR (400MHz, CHLOROFORM-d) δ = 4.32 (br s, 1H), 3.71 - 3.60 (m, 1H), 1.93 (br s, 6H), 1.88 (br d, J=9.3 Hz, 6H), 1.05 (d, J=6.5 Hz, 6H). ESI [M+H] =256.0
Preparation of compound 162.
Figure imgf000119_0002
161 162
Genneral method N, (l-carbamoyl-4-bicyclo[2.2.2]octanyl) N-isopropylcarbamate . 1H NMR (400MHz, CHLOROFORM-d) δ = 5.56 - 5.31 (m, 2H), 4.34 (br s, 1H), 3.72 - 3.57 (m, 1H), 1.96 (br s, 6H), 1.97 - 1.93 (m, 1H), 1.92 - 1.80 (m, 6H), 1.05 (d, J=6.6 Hz, 6H). ESI [M+H] =255.3
Preparation of compound 163.
Figure imgf000119_0003
162 163
General method L, (l-carbamothioyl-4-bicyclo[2.2.2]octanyl) N-isopropylcarbamate. ESI [M+H] =271.3 Preparation of compound 164.
Figure imgf000120_0001
163 164
General method M, (l-thiazol-2-yl-4-bicyclo[2.2.2]octanyl) N-isopropylcarbamate. ESI [M+H] =295.0
Preparation of compound 165
Figure imgf000120_0002
164 165
General method J, [1 -(5-bromothiazol-2-yl)-4-bicyclo[2.2.2]octanyl] N-isopropylcarbamate. ESI [M+H] =372.8/374.8
Preparation of Ex. 61.
Figure imgf000120_0003
Ex. 61
General method K, [l-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl] thiazol-2-yl]-4-bicyclo[2.2.2]octanyl] N-isopropylcarbamate. 1H NMR (400MHz,
METHANOL-d4) δ = 8.26 (d, J=2.2 Hz, 1H), 7.74 - 7.68 (m, 2H), 7.38 - 7.32 (m, 4H), 7.30 7.23 (m, 1H), 4.43 (s, 2H), 3.71 - 3.61 (m, 1H), 2.18 (s, 12H), 1.16 - 1.09 (m, 15H). ESI [M+H] =654.3 Example 62 Synthesis of 4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(pyridin-2- ylmethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-l-yl isopropylcarbamate.
The following compound was synthesized via same method by the key intermediate 165.
Figure imgf000121_0001
Ex. 62
1H NMR (400MHz, METHANOL-d4) δ = 8.77 (br s, 1H), 8.62 - 8.52 (m, 1H), 8.34 (br s, 1H), 8.12 - 8.03 (m, 1H), 7.96 (br d, J=5.5 Hz, 1H), 7.79 - 7.66 (m, 2H), 7.43 - 7.35 (m, 1H), 4.78 (br s, 2H), 3.67 (br dd, J=6.2, 13.0 Hz, 1H), 2.19 (br s, 12H), 1.17 - 1.09 (m, 15H). ESI [M/2+H] = 328.2
Example 63 Synthesis of trans-isopropyl N-[6-[5-[2-(tert-butylsulfamoyl)-4- (isopropoxycarbonylamino)phenyl]thiazol-2-yl]-3-piperidyl]carbamate
Scheme 23:
Figure imgf000121_0002
174 175
Figure imgf000122_0001
178 Ex. 63
General method I for preparation of compound 167.
Figure imgf000122_0002
166 167
To a solution of 5-hydroxypyridine-2-carboxylic acid (60 g, 431 mmol, 1 eq.) in AcOH (200 mL)/H20 (600 mL) was added wet Pd/C (2 g, 10% content) under N2. The suspension was degassed under vacuum and purged with H2 several times. The mixture was stirred under H2 (50 psi) at 50°C for 12 hrs, then filtered and concentrated to give 5-hydroxypiperidine-2- carboxylic acid (62.61 g, crude) as a yellow oil. ESI [M+H] =146.5
Preparation of compound 168
Figure imgf000122_0003
167 168
To a mixture of 5-hydroxypiperidine-2-carboxylic acid (62 g, 427.13 mmol, 1
eq.) and Boc20 (102.54 g, 469.84 mmol, 107.94 mL, 1.1 eq.) in dioxane (500 mL) was added NaOH (34.17 g, 854.25 mmol, 2 eq.) and the mixture was stirred at 25 °C for 18 hrs. The mixture was then concentrated to remove dioxane and the pH was adjusted to 2-3 by addition of IN HC1 solution. The aqueous phase was extracted with 2-Me-THF (500 mL * 3). The combined organic layers was dried over Na2S04, filtered and concentrated to give 1- tert-butoxycarbonyl-5- ydroxy- piperidine-2-carboxylic acid (40 g, crude) as yellow oil. ESI [M+Na] =267.9 Preparation of compound 169
Figure imgf000123_0001
168
169
General method A, O -benzyl Oi-tert-butyl 5-hydroxypiperidine-l,2-dicarboxylate . ESI [M+Na+] =358.0
Preparation of compound 170
Figure imgf000123_0002
169 170
A mixture of O -benzyl 01-tert-butyl 5-hydroxypiperidine-l,2-dicarboxylate (10 g, 29.82 mmol, 1 eq.), TEA (3.62 g, 35.78 mmol, 4.98 mL, 1.2 eq.) in DCM (100 mL) was added methanesulfonyl chloride (4.10 g, 35.78 mmol, 2.77 mL, 1.2 eq.) at 0 °C and the mixture was stirred at 25 °C 1 hr. The mixture was then washed with H20 (50 mL) and the organic layer was dried and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether/Ethyl acetate=40/l to 5: 1) to afford Oi-benzyl Ol-tert-butyl 5- methylsulfonyloxypiperidine-l,2-dicarboxylate (9.5 g, 18.98 mmol, 63.65% yield, 82.6% purity) as yellow oil.
A mixture of Oi-benzyl Ol-tert-butyl 5-methylsulfonyloxypiperidine-l,2- dicarboxylate (9.5 g, 22.98 mmol, 1 eq.), NaN3 (8.96 g, 137.85 mmol, 6 eq.) in DMF (20 mL) was stirred at 100°C for 4 hrs. Then the mixture was quenched with sat.aq.Na2S03 (30 mL) and extracted with EtOAc (100 mL * 3). The combined organic layers were washed with brine (40 mL * 3), dried over Na2S04, filtered and concentrated to give Oi-benzyl Oi-tert-butyl 5- azidopiperidine-l,2-dicarboxylate (8.82 g, crude) as yellow oil . Preparation of compound 171
Figure imgf000124_0001
A mixture of 02-benzyl Oi-tert-butyl 5-azidopiperidine-l,2-dicarboxylate (8 g, 22.20 mmol, 1 eq.), triphenylphosphane (8.73 g, 33.30 mmol, 1.5 eq.), in H20 (50 mL) and THF (50 mL) was stirred at 45 °C for 4 hrs. The mixture was concentrated to remove the THF and pH was adjust to 2-3 by addition of IN HC1 solution. The aqueous phase was extracted with MTBE (30 mL) and then aqueous phase was basified to adjust pH to 9 and extracted with EtOAc (50 mL * 3). The combined organic layers were dried over Na2S04 filtered and concentrated to give 02-benzyl Ol-tert-butyl 5-aminopiperidine- 1 ,2-dicarboxylate (6 g, crude) as yellow oil. ESI [M+H]=335.2
Preparation of compound 172
Figure imgf000124_0002
171 172
General method D, 02-benzyl Oi-tert-butyl 5-(isopropoxycarbonylamino)piperidine- 1,2- dicarboxylate. ESI [M+H] =421.2
Preparation of compound 173
Figure imgf000124_0003
172 173
General method O, l-tert-butoxycarbonyl-5-isopropoxycarbonyloxy-piperidine-2- carboxylic acid. ESI [M+H] =331.2 Preparation of compound 174
Figure imgf000125_0001
173 174
A mixture of l-tert-butoxycarbonyl-5-(isopropoxycarbonylamino)piperidine- 2-carboxylic acid (1.3 g, 3.93 mmol, 1 eq.), NH4C1 (315.73 mg, 5.90 mmol, 1.5 eq.), TEA (1.19 g, 11.80 mmol, 1.64 mL, 3 eq.) and HBTU (1.64 g, 4.33 mmol, 1.1 eg.) in ACN (10 mL) as stirred at 25°C for 2 hrs and then concentrated. The mixture was then poured into H20 (20 mL) and extrated with EtOAc (20ml*3). The combined organic layers were dried over Na2S04, filtered and concentrated. The mixture was purified by prep-HPLC(column: Phenomenex Gemini C18 250*50 10u;mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN] ;B%: 10%-40%,20min) to give tert-butyl 2-carbatnoyl-5- (isopropoxycarbonylamino)piperidine- 1-carboxylate (1 g, 3.01 mmol, 76.38% yield, 99% purity) as a white solid. ESI [M+H] =330.2
Preparation of compound 175
Figure imgf000125_0002
174 175
General method L, trans-tert-butyl 2-carbamothioyl-5-(isopropoxycarbonylamino) piperidine- 1 -carboxylate . ESI [M+H] =346.1
Preparation of compound 176
Figure imgf000125_0003
To a solution of trans-tert-butyl 2-carbamothioyl-5-(isopropoxycarbonylamino) piperidine- 1-carboxylate (650 mg, 1.88 mmol, 1.0 eq.) in tol. (20 mL) were added BUFFER (784.76 mg, 2.82 mmol, 1.5 eq.) and 2-chloroacetaldehyde (3.69 g, 18.82 mmol, 3.03 mL, 10 eq.). The mixture was stirred at 100°C for 1.5 hrs and then poured into water (20 mL) and extracted with EtOAc (10 mL x 3). The combined organic phase was dried over Na2S04, filtered and concentrated. The residue was purified by basic prep-HPLC to give trans-tert- butyl 5- (isopropoxycarbonylamino)-2-thiazol-2-yl-piperidine-l-carboxylate (80 mg, 216.52 umol, 11.51% yield) as a yellow solid. 1H NMR (400MHz, METHANOL-d4) δ = 7.75 (d, J=3.4 Hz, 1H), 7.58 (d, J=2.9 Hz, 1H), 5.60 (br s, 1H), 4.95 - 4.90 (m, 1H), 4.23 (br d, J=14.7 Hz, 1H), 3.68 (br s, 1H), 3.11 - 2.94 (m, 1H), 2.42 - 2.31 (m, 1H), 2.30 - 2.17 (m, 1H), 1.93 - 1.79 (m, 1H), 1.75 - 1.64 (m, 1H), 1.50 (s, 9H), 1.30 - 1.23 (m, 6H)
Preparation of compound 177
Figure imgf000126_0001
General method J, trans-tert-butyl 2-(5-bromothiazol-2-yl)-5-(isopropoxycarbonyl amino)piperidine-l-carboxylate. ESI [M+H] =450.2/448.2
Preparation of compound 178
Figure imgf000126_0002
178
General method B, trans-tert-butyl 2-[5-[2-(tert-butylsulfamoyl)-4-
(isopropoxycarbonylamino)phenyl]thiazol-2-yl]-5-(isopropoxycarbonylamino)piperidine-l- carboxylate. ESI [M+H] =682.3
Preparation of Ex. 63
Figure imgf000126_0003
178 Ex. 63 General method C, trans-isopropyl N-[6-[5-[2-(tert-butylsulfamoyl)-4- (isopropoxycarbonylamino)phenyl]thiazol-2-yl]-3-piperidyl]carbamate. 1H NMR
(400MHz, METHANOL-d4) δ = 8.40 (d, J=2.0 Hz, 1H), 7.86 (s, 1H), 7.65 (dd, J=2.2, 8.4 Hz, 1H), 7.38 (d, J=8.4 Hz, 1H), 5.05 - 4.94 (m, 1H), 4.85 - 4.81 (m, 1H), 4.67 (dd, J=2.9, 11.9 Hz, 1H), 3.92 - 3.82 (m, 1H), 3.60 (br dd, J=3.6, 11.8 Hz, 1H), 2.92 (t, J=11.9 Hz, 1H), 2.48 (br dd, J=3.2, 14.2 Hz, 1H), 2.22 - 2.03 (m, 2H), 1.82 - 1.70 (m, 1H), 1.32 (d, J=6.4 Hz, 6H), 1.24 (br d, J=6.2 Hz, 6H), 1.19 (s, 9H). ESI [M+H] =582.2
Example 64 Synthesis of isopropyl ((3R,6S)-6-(5-(4-(3-benzylureido)-2-(N-(tert- butyl)sulfamoyl)phenyl)thiazol-2-yl)piperidin-3-yl)carbamate.
The following compound was synthesized via same method by the key intermediate 178.
Figure imgf000127_0001
Ex. 64
1H NMR (400MHz, METHANOL-d4) δ = 8.30 (d, J=2.2 Hz, 1H), 7.86 (s, 1H), 7.66 (dd, J=2.2, 8.4 Hz, 1H), 7.38 - 7.20 (m, 6H), 4.85 - 4.82 (m, 1H), 4.70 (dd, J=3.2, 12.0 Hz, 1H), 4.42 (s, 2H), 3.93 - 3.82 (m, 1H), 3.62 (br dd, J=3.5, 11.7 Hz, 1H), 3.00 - 2.90 (m, 1H), 2.52 - 2.44 (m, 1H), 2.23 - 2.04 (m, 2H), 1.84 - 1.70 (m, 1H), 1.26 - 1.21 (m, 6H), 1.18 (s, 9H). ESI [M+H] =629.2
Example 65 Synthesis of isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[3-(isopropoxycarbonyl amino)cyclobutyl]thiazol-5-yl]phenyl]carbamate
Scheme 24:
Figure imgf000127_0002
179 180 181
Figure imgf000127_0003
Figure imgf000128_0001
Figure imgf000128_0002
Preparation of compound 180 ethyl chloroformate
HOOC- , , /Λ NH3 H2O ^ ¾
Y *NHBoc THF/dioxane H2N "γ NHBoc
179 180
To a solution of trans-3-(tert-butoxycarbonylamino)cyclobutanecarboxylic acid (500 mg, 2.32 mmol, 1 eq.), DIEA (750.54 mg, 5.81 mmol, 1.01 mL, 2.5 eq.) in THF (10 mL) was added ETHYL CHLOROFORMATE (277.29 mg, 2.56 mmol, 1.1 eq.) and the mixture was stirred 0°C for lhr. Then it was added into NH3.H20 (1.30 g, 9.29 mmol, 1.43 mL, 25% purity, 4 eq.) in THF (10 mL) and dioxane (10 mL) and the mixture was stirred at 25 °C for 1 hr. The mixture was washed with IN HC1 (20 mL), sat.aq.Na2C03 (20 mL) and the organic phase was dried over Na2S04, filtered and concentrated to give trans-tert-butyl N-(3- carbamoylcyclobutyl) carbamate (0.39 g, crude) as a white solid. 1H NMR (400MHz, DMSO-d6) δ = 7.20 - 7.07 (m, 2H), 6.71 (br s, IH), 4.09 - 4.00 (m, IH), 2.72 (br t, J=9.3 Hz, IH), 2.23 (br t, J=8.7 Hz, 2H), 2.01 (q, J=9.8 Hz, 2H), 1.34 (s, 9H. ESI [M+Na]=237.1
Preparation of compound 181
Figure imgf000128_0003
180 181
General method L, trans-tert-butyl N-(3-carbamothioylcyclobutyl)carbamate. IH NMR (400MHz, DMSO-d6) δ = 9.33 (br s, IH), 9.02 (br s, IH), 7.18 (br d, J=5.5 Hz, IH), 4.16 - 4.03 (m, IH), 3.33 (br s, IH), 2.45 (br d, J=12.2 Hz, 2H), 2.15 (br d, J=6.5 Hz, 2H), 1.37 (br s, 9H) Preparation of compound 182
Figure imgf000129_0001
181 182
General method M, trans-tert-butyl N-(3-thiazol-2-ylcyclobutyl)carbamate. 1H NMR (400MHz, CHLOROFORM-d) δ = 7.62 (d, J=3.2 Hz, 1H), 7.14 (dd, J=3.3, 8.3 Hz, 1H), 4.79 (br s, 1H), 4.34 (br s, 1H), 3.47 - 3.36 (m, 1H), 2.81 (br d, J=8.8 Hz, 1H), 2.66 (ddd, J=4.5, 7.9, 12.8 Hz, 1H), 2.46 - 2.34 (m, 1H), 2.17 - 2.09 (m, 1H), 1.38 (s, 9H). ESI[M+H]=255.3
Preparation of compound 183
Figure imgf000129_0002
182 183
General method J, trans-tert-butyl N-[3-(5-bromothiazol-2-yl)cyclobutyl]carbamate.
ESI[M+H]=335.1/333.1
Preparation of compound 184
Pd(PPh3)4, Na2C03, KF
Figure imgf000129_0003
183 Tol./EtOH/H20, 80°C
184
General method K, trans-tert-butyl N-[3-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl] thiazol-2-yl]cyclobutyl]carbamate. ESI [M+H]=481.0
Preparation of compound 185
Figure imgf000129_0004
184 185 General method F, trans-5-amino-2-[2-(3-aminocyclobutyl)thiazol-5-yl]-N-tert- butyl- benzenesulfonamide. ESI [M+H] =381.2
Preparation of Ex. 65
Figure imgf000130_0001
General method D, isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[3-
(isopropoxycarbonylamino)cyclobutyl]thiazol-5-yl]phenyl]carbamate. 1H NMR (400MHz, METHANOL-d4) δ = 8.34 (d, J=1.5 Hz, 1H), 7.76 - 7.65 (m, 2H), 7.38 (t, J=8.5 Hz, 1H), 5.04 - 4.94 (m, 1H), 4.82 (br d, J=6.4 Hz, 1H), 4.42 - 4.32 (m, 0.5H), 4.13 (br t, J=8.0 Hz, 0.5H), 3.87 - 3.78 (m, 0.5H), 3.58 - 3.48 (m, 0.5H), 2.82 (dq, J=2.6, 7.9 Hz, 1H), 2.72 - 2.64 (m, 1H), 2.60 - 2.50 (m, 1H), 2.34 - 2.23 (m, 1H), 1.31 (d, J=6.2 Hz, 6H), 1.22 (br d, J=6.0 Hz, 6H), 1.13 (s, 9H). ESI [M+H] =553.4
Examples 66A and 66B
Scheme 25:
Figure imgf000130_0002
37
Figure imgf000131_0001
Figure imgf000131_0002
Figure imgf000131_0003
Preparation of compound 31.
Figure imgf000131_0004
To a solution of 2-bromobenzenesulfonyl chloride (100.00 g, 391.36 mmol, 1.00 eq.) in H2S04 (1.0 L) was added a solution of HN03 (79.95 g, 1.21 mol, 57.11 mL, 95% purity, 3.08 eq.) in H2S04 (0.50 L) drop-wise at 0°C. The mixture was stirred at 26°C for 2 hrs. TLC (Petroleum ether: EtOAc =10: 1, Rf=0.40) showed the reaction was complete. The mixture was added slowly to ice water (5 L) with vigorous stirring and then filtered. The filter cake was washed with H20 (1 L X 3) and dried to give 2-bromo-5-nitro-benzenesulfonyl chloride (105.00 g, crude) as a yellow solid. 1H NMR (400MHz, CHLOROFORM-d) δ = 8.98 (d, J=2.4 Hz, 1H), 8.37 (dd, J=2.5, 8.7 Hz, 1H), 8.09 (d, J=8.6 Hz, 1H).
Preparation of compound 32.
Figure imgf000131_0005
32 To a mixture of 2-methylpropan-2-amine (200 g, 2.73 mol, 287.36 mL, 3.29 eq.) and DMAP (10 g, 81.85 mmol, 0.1 eq.) in DCM (2 L) was added 2-bromo-5-nitro-benzenesulfonyl chloride (250 g, 831.91 mmol, 1 eq.) portionwise at 0°C. The mixture was warmed to 15°C and stirred for 1 hr. LCMS showed the reaction was complete. The mixture was washed with HC1 (1 N, 2 L), sat.aq. NaHC03 (500 mL) and brine (500 mL), dried over Na2S04, filtered and concentrated to give 2-bromo-N-tert-butyl-5-nitro-benzenesulfonamide (240 g, crude) as a gray solid. ESI [M+H] = 336.9/338.9.
Preparation of compound 33.
Figure imgf000132_0001
32 33
To a solution oi2-bromo-N-tert-butyl-5-nitro-benzenesulfonamide (35 g, 103.80 mmol, 1 eq.) in DMA (300 mL), were added thiazole (26.51 g, 311.40 mmol, 3 eq.), Pd(OAc)2 (2.33 g, 10.38 mmol, 0.1 eq.) and KOAc (30.56 g, 311.40 mmol, 3 eq.). The mixture was stirred at 140°C for 16 hrs under N2. LCMS showed the reaction was complete, the mixture was poured into water (3 L) and filtered. The filter cake was washed with water (200 mL x 3) and then dried to give N-tert-butyl-5-nitro-2-thiazol-5-yl-benzenesulfonamide (25.1 g, crude) as a black brown solid. 1H NMR (400MHz, DMSO-d6) δ = 9.30 (s, 1H), 8.81 (d, J=2.4 Hz, 1H), 8.46 (dd, J=2.4, 8.3 Hz, 1H), 8.12 (s, 1H), 7.83 (d, J=8.3 Hz, 1H), 7.60 (s, 1H), 1.07 (s, 9H). ESI [M+H] = 342.0.
Preparation of compound 34.
Figure imgf000132_0002
To a solution oiN-tert-butyl-5-nitro-2-thiazol-5-yl-benzenesulfonamide (15 g, 43.94 mmol, 1 eq.) in AcOH (200 mL), were added KOAc (21.56 g, 219.68 mmol, 5 eq.) and Br2 (35.11 g, 219.68 mmol, 5 eq.). The mixture was stirred at 80°C for 3 hrs. LCMS showed the reaction was complete, the mixture was quenched by sat.aq.Na2C03 (1 L) and extracted with EtOAc (300 mL x 3). The combined organic phase was dried over Na2S04, filtered and concentrated to give 2-(2-bromothiazol-5-yl)-N-tert-butyl-5-nitro-benzenesulfonamide (18 g, crude) as a green solid which was used without any purification.
Preparation of compound 35.
Figure imgf000133_0001
34 35
To a solution of 2-(2-bromothiazol-5-yl)-N-tert-butyl-5-nitro-benzenesulfonamide (8.5 g, 20.22 mmol, 1 eq.) in EtOH (70 mL), THF (40 mL) and H20 (20 mL), were added Fe (3.39 g, 60.67 mmol, 3 eq.) and NH4C1 (3.25 g, 60.67 mmol, 2.12 mL, 3 eq.). The mixture was stirred at 90°C for 1 hr. LCMS showed the reaction was complete. The mixture was filtered, the filtrate was concentrated to remove organic solvent and extracted with DCM (200 mL x 2). The organic phase was dried over Na2S04, filtered and concentrated to afford 5-amino-2- (2-bromothiazol-5-yl)-N-tert-butyl- benzenesulfonamide (6 g, crude) as a yellow solid. 1HNMR showed the structure was correct. IH NMR (400MHz, DMSO-d6) δ = 7.58 - 7.52 (m, IH), 7.30 (d, J=2.4 Hz, IH), 7.11 (d, J=8.3 Hz, IH), 6.99 (s, IH), 6.73 (dd, J=2.4, 8.3 Hz, IH), 5.94 (s, 2H), 1.07 (s, 9H). ESI [M+H] = 390.0/392.0.
Preparation of compound 36.
Figure imgf000133_0002
35 36
General method D, (4-nitrophenyl) N-[4-(2-bromothiazol-5-yl)-3-(tert- butylsulfamoyl)pheny I] carbamate as DCM solution ESI [M+H] = 555.0/557.0
Preparation of compound 37.
Figure imgf000133_0003
36 37 General method H, l-benzyl-3-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl) phenyljurea. ESI [M+H] = 523.0/525.0.
Preparation of compound 38.
Figure imgf000134_0001
37 38
General method B {Suzuki reaction), tert-butyl-N-[4-[5-[4-(benzylcarbamoylamino)-2- (tert- butylsulfamoyl)phenyl]thiazol-2-yl]cyclohex-3-en-l-yl]carbamate ( Compound 38).
To a solution of Compound 17 (1 eq.) in dioxane and H20, were added Pd(dppf)Cl2 (0.1 eq.), Comound 37 (0.9 eq.) and Na2C03 (3 eq.). The mixture was stirred at 80°C for 12 hrs under N2. LCMS showed the reaction was complete. The mixture was concentrated and the residue was purified by prep-TLC (PE/EtOAc=l : l) to yield product 38. ESI [M+H] = 640.5
Preparation of compound 39.
Figure imgf000134_0002
tert-butyl N-[4-[5-[4-(benzylcarbamoylamino)-2-(tert- butylsulfamoyl)phenyl]thiazol-2- yljcyclohexyljcarbamate .
To a solution of Compound 38 (1 eq.) and AcOH (0.1 eq.) in EtOAc was added Pd/C (10% purity, 1.00 eq.). The mixture was stirred under H2 (15 psi) at 40°C for 1 hr. LCMS showed the reaction was complete and then the mixture was filtered and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether/Ethyl acetate =10/1 to 1 : 1) to give Compound 39. ESI [M+H] = 642.5
Preparation of compound 40.
Figure imgf000134_0003
General method F, l-[4-[2-(4-aminocyclohexyl)thiazol-5-yl]-3-(tert-butylsulfamoyl) phenyl]-3-benzyl-urea. ESI [M+H] = 542.5.
Preparation of compound 41
Figure imgf000135_0001
General method D, isopropyl N-[4-[5-[4-(benzylcarbamoylamino)-2-(tert- butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate, ESI [M+H] = 628.3.
Preparation of Compound Ex. 66A and Ex. 66B.
Figure imgf000135_0002
Ex. 66B
Compound 41 was separated by SFC (Instrument: Thar SFC80 preparative SFC; Column: Chiralpak AD-H 250*30mm i.d. 5u; Mobile phase: A for C02 and B for IPA(0.1%NH3H2O); Gradient: B%=42%; Flow rate:70 g/min; Wavelength:220 nm; Column temperature: 40 °C ; System back pressure: 100 bar; Cycle time:20 min; Injection amount: 4 mg per injection), and then purified by prep-HPLC (Column: Luna C18 100*30 5u; mobile phase:
[water(0.1%TFA)-ACN];B%: 35%-75%,5min ). trans-isopropyl N-[4-[5-[4- (benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate Ex. 66A (11.54 mg, 18.38 umol, 5.53% yield, 100% purity) and cis-isopropyl N-[4-[5-[4- (benzylcarbamoylamino )-2-(tert-butylsulfamoyl)phenyl] thiazol-2-yl]cyclohexyl]carbamate Ex. 66B (9.38 mg, 14.94 umol, 4.50% yield, 100% purity) were obtained. Trans-isopropyl N-[4-[5-[4-(benzylcarbamoylamino )-2-( tert-butylsulfamoyl)phenyl] thiazol-2-yl]cyclohexyl]carbamate, 1H NMR (400MHz, METHANOL-d4) δ = 8.24 (d, J=2.2 Hz, 1H), 7.72 (s, 1H), 7.69 (dd, J=2.6, 8.3 Hz, 1H), 7.37 - 7.30 (m, 5H), 7.27 - 7.22 (m, 1H), 4.85 - 4.78 (m, 1H), 4.41 (s, 2H), 3.50 - 3.39 (m, 1H), 3.06 - 2.96 (m, 1H), 2.22 (br d, J=11.8 Hz, 2H), 2.07 (br d, J=10.1 Hz, 2H), 1.76 - 1.62 (m, 2H), 1.41 (dq, J=3.1, 12.7 Hz, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.11 (s, 9H). ESI [M+H] = 628.2.
Cis-isopropyl N-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl
]thiazol-2-yl]cyclohexyl]carbamate. 1H NMR (400MHz, METHANOL-d4) δ = 8.25 (d, J=2.2 Hz, 1H), 7.74 (s, 1H), 7.70 (dd, J=2.2, 8.3 Hz, 1H), 7.38 - 7.30 (m, 5H), 7.28 - 7.22 (m, 1H), 4.82 (br d, J=6.1 Hz, 1H), 4.41 (s, 2H), 3.74 (br s, 1H), 3.20 - 3.13 (m, 1H), 2.03 - 1.95 (m, 4H), 1.86 - 1.72 (m, 4H), 1.23 (d, J=6.1 Hz, 6H), 1.12 (s, 9H). ESI [M+H] = 628.2.
Examples 67A and 67B
Scheme 26:
Figure imgf000136_0001
Ex. 67A Ex. 67B Preparation of compound 42.
Figure imgf000137_0001
35
General method D, isopropyl N-[4-(2-bromothiazol-5-yl) -3-(tert-butylsulfamoyl) phenyljcarbamate. ESI [M+H] =476.0/478.0.
Preparation of compound 43
Figure imgf000137_0002
General method B, isopropyl N-[4-[2-[4-(tert -butoxycarbonylamino)cyclohexen-l-yl] thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate. ESI [M+H] =593.3.
Preparation of compound 44.
Figure imgf000137_0003
43
General method I, isopropyl N-[4-[2-[4-(tert-butoxycarbonylamino)cyclohexyl] thiazol-5- yl]-3-(tert-butylsulfamoyl)phenyl]carbamate. ESI [M+H] =595.3.
Preparation of compound 45.
Figure imgf000137_0004
44
General method F, isopropyl N-[4-[2-(4-aminocyclohexyl) thiazol-5-yl]-3-(tert- butylsulfamoyl)phenyl] carbamate . ESI [M+H] =495.2. Preparation of compound 46.
Figure imgf000138_0001
General method D, isopropyl N-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino) cyclohexyl]thiazol-5-yl]phenyl]carbamate. ESI [M+H] =581.2.
Preparation of Ex. 67 A and Ex. 67B.
Figure imgf000138_0002
Ex. 67B
Compound 46 was separated by SFC (Instrument: Thar SFC80 preparative SFC; Column: ChiralpakAD-H 250*30mm i.d. 5u; Mobile phase: A for C02 and B for IPA(0.1%NH3H2O); Gradient: B%=30%; Flow rate:70 g/min; Wavelength:220 nm; Column temperature: 40°C; System back pressure: 100 bar; Cycle time:8 min; Injection amount: 3 mg per injection); and then purified by prep-HPLC (Column: Agela Durashell C18 150*25 5u; mobile phase:
[water(0.1%TFA)-ACN];B%: 55%-85%,12min), trans-isopropylN-[3-(tert-butylsulfamoyl)- 4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate Ex. 67 (5.76 mg, 100% purity) and cis-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4- (isopropoxycarbonylamino) cyclohexyl]thiazol-5-yl]phenyl]carbamate Ex. 67B (3.95 mg , 100% purity) were obtained as a pale yellow solid.
Trans-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl] thiazol-5-yl]phenyl]carbamate (Compound SI 2). 1H NMR (400MHz, METHANOL-d4) δ = 8.37 (d, J=2.3 Hz, 1H), 7.75 (s, 1H), 7.69 (dd, J=2.2, 8.4 Hz, 1H), 7.40 (d, J=8.3 Hz, 1H), 5.01 (td, J=6.3, 12.5 Hz, 1H), 4.86 - 4.82 (m, 1H), 3.52 - 3.42 (m, 1H), 3.04 (tt, J=3.5, 12.0 Hz, 1H), 2.35 - 2.19 (m, 2H), 2.15 - 1.99 (m, 2H), 1.72 (dq, J=3.0, 12.9 Hz, 2H), 1.43 (dq, J=3.3, 12.6 Hz, 2H), 1.34 (d, J=6.2 Hz, 6H), 1.25 (br d, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H] = 581.2.
Cis-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-
(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate ( Compound S13). 1H
NMR (400MHz, METHANOL-d4) δ = 8.37 (d, J=2.2 Hz, IH), 7.76 (s, IH), 7.70 (dd, J=2.2, 8.3 Hz, IH), 7.40 (d, J=8.3 Hz, IH), 5.01 (td, J=6.3, 12.5 Hz, IH), 4.84 (br s, IH), 3.77 (br s, IH), 3.23 - 3.15 (m, IH), 2.05 - 1.98 (m, 4H), 1.89 - 1.72 (m, 4H), 1.34 (d, J=6.2 Hz, 6H), 1.25 (d, J=6.1 Hz, 6H), 1.16 (s, 9H). ESI [M+H] = 581.2
Example 68 Synthesis of isopropyl N-[l-[5-[4-(benzylcarbamoylamino)-2-(tert-butyl sulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl]carbamate.
Scheme 27:
Figure imgf000139_0001
19
Preparation of compound 24.
Figure imgf000139_0002
19 24
A mixture of l-benzyl-3-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl)phenyl]urea (0.06 g, 114.62 umol, 1.0 eq.), tert-butyl N-(4-piperidyl)carbamate (40 mg, 199.72 umol, 1.74 eq.), Xantphos (6.63 mg, 11.46 umol, 0.1 eq.), Pd(OAc)2 (2.57 mg, 11.46 umol, 0.1 eq.) and Cs2C03 (112.04 mg, 343.86 umol, 3.0 eq.) in ACN (3 mL) was heated to 80°C for 12 hrs under N2. LCMS showed the reaction was complete and the mixture was concentrated. The residue was purified by prep-TLC (Petroleum ether: Ethyl acetate = 1: 1) to give tert-butyl N-[l-[5-[4-(benzylcarbamoylamino) -2-(tert- butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl]carbamate (0.03 g, crude) as a yellow solid. ESI [M+H] = 643.1.
Preparation of compound 25.
Figure imgf000140_0001
Tert-butyl N-[l-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl] thiazol-2-yl]- 4-piperidyl]carbamate (30 mg, 46.67 umol, 1.0 eq.) was dissolved into HCl/MeOH (4M, 1 mL) and the mixture was stirred at 20°C for 0.5 hr. LCMS showed the reaction was complete and the mixture was concentrated to give l-[4-[2-(4-amino-l-piperidyl)thiazol-5-yl]-3-(tert- butylsulfamoyl)phenyl]-3-benzyl-urea (27 mg, crude, HC1 salt) as a yellow solid. ESI
[M+H] = 543.2.
Preparation of Ex. 68.
Figure imgf000140_0002
25 Ex. 68
General method D, isopropyl N-[l-[5-[4-(benzylcarbamoylamino)-2-(tert-butyl
sulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl]carbamate. 1H NMR (400MHz, METHANOL- d4) δ = 8.24 (d, J=2.2 Hz, 1H), 7.69 (dd, J=2.4, 8.4 Hz, 1H), 7.39 (d, J=8.4 Hz, 1H), 7.36 - 7.19 (m, 6H), 4.82 - 4.73 (m, 1H), 4.40 (s, 2H), 3.92 (br d, J=13.7 Hz, 2H), 3.75 - 3.68 (m, 1H), 3.43 (br t, J=11.0 Hz, 2H), 2.10 - 2.02 (m, 2H), 1.71 - 1.60 (m, 2H), 1.22 (br d, J=6.2 Hz, 6H), 1.18 (s, 9H). ESI [M+H] = 629.2. Example 69 Synthesis of isopropyl N-[l-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxy carbonylamino)phenyl]thiazol-2-yl]-4-piperidyl]carbamate.
Scheme 28:
Figure imgf000141_0001
Example 69
Preparation of compound 26.
Figure imgf000141_0002
17 26
General method D, isopropyl N-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl) phenyljcarbamate. ESI [M+H] = 478.0/476.0.
Preparation of compound 27.
Figure imgf000141_0003
26
To a solution of isopropyl N-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl)
phenyljcarbamate (0.05 g, 104.95 umol, 1.0 eq.) in MeCN (2 mL) were added Cs2CC>3 (102.59 mg, 314.86 umol, 3.0 eq.), KI (17.42 mg, 104.95 umol, 1.0 eq.) and tert-butyl N-(4- piperidyl)carbamate (105.10 mg, 524.76 umol, 5.0 eq.). The mixture was stirred at 95°C for 12 hrs and then concentrated. The residue was purified by prep-TLC (Si02, Petroleum ether: Ethyl acetate = 4:3) to give N-[4-[2-[4-(tert-butoxycarbonylamino)-l-piperidyl]thiazol-5-yl]- 3-(tert-butylsulfamoyl) phenyl] carbamate (0.025 g, crude) as a white solid. ESI [M+H] = 596.2.
Preparation of compound 28.
Figure imgf000142_0001
Isopropyl N-[4-[2-[4-( tert-butoxycarbonylamino )-l-piperidyl]thiazol-5-yl]-3-(tert- butylsulfamoyl)pheny I] carbamate (0.025 g, 41.96 umol, 1.0 eq.) was dissolved
into HCl/MeOH (4M, 1.5 mL) and the mixture was stirred at 20°C for 0.5 hr. LCMS showed the reaction was complete and the mixture was concentrated to give the N-[4-[2-(4-amino-l- piperidyl)thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl] carbamate (0.02 g, crude, HC1 salt) as a white solid. ESI [M+H] = 496.2.
Preparation of Ex. 69.
Figure imgf000142_0002
28 Ex. 69
General method D, isopropyl N-[l-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxy
carbonylamino)phenyl]thiazol-2-yl]-4-piperidyl]carbamate. 1H NMR (400MHz,
METHANOL-d4) δ = 8.35 (s, 1H), 7.66 (br d, J=8.4 Hz, 1H), 7.42 (d, J=8.4 Hz, 1H), 7.31 (s, 1H), 4.98 (td, J=6.2, 12.5 Hz, 1H), 4.84 - 4.77 (m, 1H), 3.93 (br d, J=13.5 Hz, 2H), 3.77 - 3.67 (m, 1H), 3.44 (br t, J=12.3 Hz, 2H), 2.06 (br dd, J=3.1, 13.2 Hz, 2H), 1.71 - 1.61 (m, 2H), 1.31 (d, J=6.4 Hz, 6H), 1.23 (br d, J=6.2 Hz, 6H), 1.19 (s, 9H). ESI [M+H] = 582.2. Example 70 Synthesis of trans-isopropyl N-[4-[5-[2-(tert-butylsulfamoyl)-4-[(2- fluorophenyl)methoxy carbonylamino]phenyl]thiazol-2-yl]cyclohexyl]carbamate.
Scheme 29:
Figure imgf000143_0001
Ex. 70
Preparation of compound 2. " 'NHBoc
Figure imgf000143_0002
To a mixture of trans-4-(tert-butoxycarbonylamino)cyclohexanecarboxylic acid (65.0 g, 267.2 mmol, 1.0 eq.), NH4C1 (21.4 g, 400.7 mmol, 1.5 eq.) and TEA (801.5 mmol, 111.6 mL, 3 eq.) in MeCN (1.3 L) was added HBTU (111.5 g, 293.9 mmol, 1.1 eq.) and the mixture was stirred at 25 °C for 3 hrs. The mixture was filtered and then the filter cake was washed with petroleum ether (200 mL) and dried to give trans-tert-butyl N-(4- carbamoylcyclohexyl)carbamate (140 g, crude, 2 batches) as a white solid. 1H NMR (METHANOL-d4, 400MHz) δ = 3.25-3.34 (m, 1H), 2.14 (tt, J=12.3, 3.5 Hz, 1H), 1.83- 1.99 (m, 4H), 1.52 (qd, J=13.1, 2.9 Hz, 2H), 1.42 (s, 9H), 1.21 (qd, J=12.7, 3.5 Hz, 2H)
Preparation of compound 3.
Figure imgf000144_0001
2 3
A mixture of trans-tert-butyl N-(4-carbamoylcyclohexyl)carbamate (90.0 g, 371.4 mmol, 1.0 eq.), Na2C03 (39.4 g, 371.4 mmol, 1.0 eq.) and Lawesson's reagent (82.6 g, 204.3 mmol, 0.55 eq.) in 2-Me-THF (600 mL) was stirred at 80°C for 2 hrs and then the reaction mixture was poured into H20 (200 mL). The aqueous phase was extracted with EtOAc (500 mL x 2). The combined organic layers were dried over Na2S04, filtered and concentrated to give trans-tert-butyl N-(4-carbamothioylcyclohexyl)carbamate (180 g, crude, 2 batches) as a white solid. 1H NMR (METHANOL-d4, 400MHz) δ = 3.35-3.46 (m, 1H), 2.87-3.00 (m, 1H), 2.09-2.20 (m, 2H), 1.99-2.09 (m, 2H), 1.54- 1.68 (m, 2H), 1.26- 1.45 (m, 2H), 1.14- 1.25 (m, 9H)
Preparation of compound 4.
Figure imgf000144_0002
3 4
A mixture of trans-tert-butyl N-(4-carbamothioylcyclohexyl)carbamate (180.0 g, 696.7 mmol, 1.0 eq.), 2-bromo- l, l-diethoxy-ethane (137.3 g, 696.7 mmol, 1.0 eq.) and TsOH.H20 (265 g, 1.4 mol, 2 eq.) in EtOH (2.0 L) was stirred at 80°C for 6 hrs. Then the mixture was cooled to RT and adjusted to PH=9 with aq.sat.Na2C03 and Boc20 (152 g, 696.7 mmol, 1 eq.) was added. The mixture was stirred at 30°C for 3 hrs, then concentrated and diluted with H20 (2 L). The mixture was extracted with EtOAc (800 mL x 3) and the combined organic layers were dried over Na2S04, filtered and concentrated. The residue was triturated with petroleum ether (1.5 L) to give trans-tert-butyl N-(4-thiazol-2-ylcyclohexyl) carbamate (70 g, 247.87 mmol, 35.58% yield) as a white solid. 1H NMR (METHANOL-d4, 400MHz) δ = 7.67 (d, J=3.1 Hz, 1H), 7.44 (d, J=3.5 Hz, 1H), 3.33-3.44 (m, 1H), 2.93-3.06 (m, 1H), 2.12- 2.21 (m, 2H), 2.00-2.09 (m, 2H), 1.57- 1.71 (m, 2H), 1.41- 1.48 (m, 9H), 1.29- 1.38 (m, 1H), 1.13- 1.28 (m, 1H). Preparation of compound 5. '" NHBoc
Figure imgf000145_0001
A mixture of trans-tert-butyl N-(4-thiazol-2-ylcyclohexyl)carbamate (68 g, 240.8 mmol, 1 eq.) and NBS (47.1 g, 264.9 mmol, 1.1 eq.) in DMF (500 mL) was stirred at 25 °C for 10 hrs and then poured into H20 (2 L) and extracted with EtOAc (500 mL x 3). The combined organic layers were washed with brine (300 mL x 5), dried over Na2S04, filtered and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether: Ethyl acetate = 20: 1 to 10: 1) to give trans-tert-butyl N-[4-(5-bromothiazol-2- yl)cyclohexyl]carbamate (78 g, crude) as a yellow solid. ESI [M+H] = 363.0/361.0
Preparation of compound 6.
Figure imgf000145_0002
A mixture of trans-tert-butyl N-[4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (50 g, 138.4 mmol, 1 eq.) in HCI/MeOH (4 M, 700 mL) was stirred at 25 °C for 0.5 hr and then concentrated to give trans-4-(5-bromothiazol-2-yl) cyclohexanamine (45 g, crude, HCl salt) as a yellow solid. ESI [M+H] = 263.0/261.0
Preparation of compound 7.
Figure imgf000145_0003
To a solution of trans-4-(5-bromothiazol-2-yl)cyclohexanamine (45 g, HCl salt, 172.3 mmol, 1 eq.), Pyridine (61.5 mmol, 69.5 mL, 5 eq.) and DMAP (4.2 g, 34.5 mmol, 0.2 eq.) in DCM (300 mL) was added isopropyl carbonochloridate (258.5 mmol, 35.9 mL, 1.5 eq.) dropwise at 0°C. The mixture was stirred at 25 °C for 0.5 hr and then washed with HCl (IN, 1 L) and sat.aq.Na2C03 (1 L). The organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether: Ethyl acetate =15: 1 to 10: 1) to give trans-isopropyl N-[4-(5-bromothiazol-2- yl)cyclohexyl]carbamate (37 g, 106.55 mmol, 61.84% yield) as a yellow solid. 1H NMR (METHANOL-d4, 400MHz) δ = 7.60 (s, IH), 4.81 (dt, J=12.2, 6.2 Hz, IH), 3.41 (tt, J=11.6, 4.0 Hz, IH), 2.88-3.00 (m, IH), 2.10-2.20 (m, 2H), 1.98-2.07 (m, 2H), 1.55- 1.68 (m, 2H), 1.30- 1.43 (m, 2H), 1.21 (br d, J=6.2 Hz, 6H).
Preparation of compound 8.
Figure imgf000146_0001
A mixture of trans-isopropyl N-[4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (16.2 g, 46.7 mmol, 1 eq.), 5-amino-N-tert-butyl-2-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2- yl)benzenesulfonamide (19.9 g, 56.1 mmol, 1.2 eq.), KF (4.1 g, 70.1 mmol, 1.5 eq.), Na2C03 (14.9 g, 140.2 mmol, 3 eq.) and Pd(PPh3)4 (1.6 g, 1.4 mmol, 0.03 eq.) in toluene (150 mL), EtOH (150 mL) and H20 (50 mL) stirred at 80°C for 6 hrs under N2 atmosphere. The reaction mixture was concentrated and the residue was diluted with H20 (100 mL) and extracted with EtOAc (100 mL x 2). The combined organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether: Ethyl acetate= 20: 1 to 1 : 1) to give trans-isopropyl N-[4-[5-[4-amino-2- (tert-butylsulfamoyl) phenyl]thiazol-2-yl]cyclohexyl]carbamate (13 g, 26.3 mmol, 56.2% yield) as a white solid. 1H NMR (400MHz, METHANOL-d4) δ = 7.65 - 7.60 (m, IH), 7.43 (d, J=2.2 Hz, IH), 7.13 (d, J=8.3 Hz, IH), 6.83 (dd, J=2.6, 8.3 Hz, IH), 4.81 (br s, IH), 3.76 - 3.71 (m, IH), 3.05 - 2.86 (m, IH), 2.20 (br d, J=12.3 Hz, 2H), 2.06 (br d, J=10.5 Hz, 2H), 1.75 - 1.60 (m, 2H), 1.47 - 1.34 (m, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.09 (s, 9H).
Preparation of compound 9.
Figure imgf000146_0002
To a solution of trans-isopropyl N-[4-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl] thiazol-2- yl]cyclohexyl]carbamate (250 mg, 505.4 umol, 1 eq.) in DCM (4 mL) were added DMAP (6.2 mg, 50.5 umol, 0.1 eq.) , Pyridine (120 mg, 1.5 mmol, 3 eq.) and (4-nitrophenyl) carbonochloridate (153 mg, 758 umol, 1.5 eq.). The mixture was stirred at 25°C for 0.5 hr and used directly for the next step. ESI [M+H] = 660.2 Preparation of compound Ex. 70.
Figure imgf000147_0001
Ex. 70
To a solution of (2-fluorophenyl)methanol (45.9 mg, 363.8 umol, 3 eq.) and DIEA (47 mg, 363.8 umol, 3 eq.) in MeCN (1 mL) was added the aboved solution (1 mL). The mixture was stirred at 80°C for 1 hr, then concentrated and the residue was purified by prep-HPLC (Column: Waters Xbridge 150*25 5u; Mobile phase: [water (10 mM NH4HC03)-ACN] ; B%: 42%-72%, 12 min) to give trans-isopropyl N-[4-[5-[2-(tert-butylsulfamoyl)-4-[(2- fluorophenyl)methoxy carbonylamino]phenyl]thiazol-2-yl]cyclohexyl]carbamate (16.34 mg, 25.26 umol, 20.83% yield, 100% purity) as a yellow solid. 1H NMR (400MHz,
METHANOL-d4) δ = 8.36 (s, 1H), 7.77 - 7.63 (m, 2H), 7.52 (t, J=7.0 Hz, 1H), 7.44 - 7.31 (m, 2H), 7.23 - 7.08 (m, 2H), 5.29 (s, 2H), 4.83 (br s, 1H), 3.46 (br d, J=11.8 Hz, 1H), 3.00 (br t, J=11.8 Hz, 1H), 2.22 (br d, J=12.7 Hz, 2H), 2.07 (br d, J=11.4 Hz, 2H), 1.76 - 1.62 (m, 2H), 1.47 - 1.35 (m, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.12 (s, 9H). ESI [M+H] = 647.2
Example 71 Synthesis of trans-[(lS)-l-phenylethyl] N-[3-(tert-butylsulfamoyl)-4-[2-[4- (isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate.
Scheme 30:
Figure imgf000147_0002
Ex. 71
Preparation of Ex. 71.
To a solution of (lS)- l-phenylethanol (29.6 mg, 242.5 umol, 2 eq.) and DIEA (47 mg, 363.8 umol, 3 eq.) in MeCN (2 mL) was added a solution of trans-(4-nitrophenyl) N-[3-(tert- butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5- yljpheny I] carbamate (80 mg, 121.25 umol, 1 eq.) in DCM (1 mL). The mixture was stirred at 80°C for 1 hr, then concentrated and the residue was purified by prep-HPLC (Column:
Nano-Micro UniSil 5- 100 C18 ULTRA 100*250mm 5um;mobile phase: [water(0.1%TFA)-
ACN] ; B%: 55%-80%, l lmin) to give trans-[(lS)-l-phenylethyl] N-[3-(tert-butylsulfamoyl)- 4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate (12.19 mg, 18.77 umol, 15.48% yield, 99% purity) as a white solid. 1H NMR (400MHz, METHANOL- d4) δ = 8.35 (br s, IH), 7.81 - 7.65 (m, 2H), 7.48 - 7.36 (m, 5H), 7.31 (br d, J=6.8 Hz, IH), 5.89 (br d, J=6.2 Hz, IH), 4.85 (br d, J=5.5 Hz, IH), 3.47 (br s, IH), 3.03 (br s, IH), 2.24 (br d, J=11.2 Hz, 2H), 2.08 (br d, J=11.0 Hz, 2H), 1.71 (q, J=11.9 Hz, 2H), 1.61 (br d, J=6.4 Hz, 3H), 1.49 - 1.37 (m, 2H), 1.24 (br d, J=5.4 Hz, 6H), 1.13 (s, 9H). ESI [M+H] = 643.2
Example 72 Synthesis of N-[3-(tert-butylsulfamoyl)-4-[2-[4- (isopropoxycarbon lamino) cyclohexy 1] thiazol- 5-yl] phenyl] carbamate.
Scheme 31:
Figure imgf000148_0001
Ex. 72
Preparation of Ex. 72.
To a solution of 2-pyridylmethanol (26.5 mg, 242.5 umol, 2 eq.) and DIEA (47 mg,
363.8umol, 3 eq.) in MeCN (2 mL) was added a solution of trans-(4-nitrophenyl) N-[3-(tert- butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5- yljpheny I] carbamate (80 mg, 121.3 umol, 1 eq.) in DCM (1 mL). The mixture was stirred at 80°C for 1 hr, then concentrated and the residue was purified by prep-HPLC (Column: Waters Xbridge 150*25 5u; Mobile phase: [water (0.04% NH3H20 + 10 mM NH4HC03)- ACN] ; B%: 35%-65%, lO min) to give trans-2-pyridylmethyl N-[3-(tert-butylsulfamoyl)-4- [2-[4-(isopropoxycarbonylamino)cyclohexyl] thiazol-5-yl]phenyl]carbamate (18.59 mg, 28.82 umol, 23.77% yield, 97.65% purity) as a pale yellow solid. 1H NMR (400MHz, METHANOL-d4) δ = 8.56 (br d, J=4.5 Hz, IH), 8.38 (d, J=1.8 Hz, IH), 7.94 - 7.87 (m, IH), 7.78 - 7.72 (m, 2H), 7.59 (br d, J=7.8 Hz, IH), 7.44 - 7.38 (m, 2H), 5.32 (s, 2H), 4.85 (td, J=5.9, 12.0 Hz, IH), 3.47 (br t, J=11.8 Hz, IH), 3.07 - 2.96 (m, IH), 2.24 (br d, J=12.3 Hz, 2H), 2.13 - 2.04 (m, 2H), 1.77 - 1.65 (m, 2H), 1.48 - 1.37 (m, 2H), 1.24 (br d, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H] = 630.2 Example 73 Synthesis of trans-isopropyl N-[4-[5-[2-(tert-butylsulfamoyl)-4-[(4- hydroxyphenyl) methylcarbamoy lamino] phenyl] thiazol-2-yl] cyclohexy 1] carbamate.
Scheme 32:
Figure imgf000149_0001
Ex. 73
Preparation of Ex. 73.
To a solution 4-(aminomethyl) phenol (74.7 mg, 606.5 umol, 3 eq.) and DIEA (78.4 mg, 606.5 umol, 3 eq.) in DCM (2 mL) was added trans-(4-nitrophenyl) N-[3-(tert- butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5- yljpheny I] carbamate (133.4 mg, 202.2 umol, 1 eq.) in DCM (2 mL). The mixture was stirred at 25 °C for 1 hr, then concentrated and the residue was purified by prep-HPLC (TFA condition) to give trans-isopropyl N-[4-[5-[2-(tert-butylsulfamoyl)-4-[(4- hydroxyphenyl)methylcarbamoylamino]phenyl]thiazol-2-yl]cyclohexyl]carbamate (23.56 mg, 35.99 umol, 17.80% yield, 98.357% purity) as a white solid. 1H NMR (400MHz, METHANOL-d4) δ = 8.24 (d, J=2.2 Hz, 1H), 7.77 (s, 1H), 7.69 (dd, J=2.2, 8.3 Hz, 1H), 7.36 (d, J=8.3 Hz, 1H), 7.16 (d, J=8.8 Hz, 2H), 6.75 (d, J=8.3 Hz, 2H), 4.85 - 4.74 (m, 1H), 4.30 (s, 2H), 3.45 (s, 1H), 3.15 - 2.97 (m, 1H), 2.23 (br d, J=12.3 Hz, 2H), 2.08 (br d, J=13.2 Hz, 2H), 1.79 - 1.63 (m, 2H), 1.48 - 1.34 (m, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.12 (s, 9H). ESI [M+H] = 644.2
Example 74 Synthesis of trans-isopropyl N-[4-[5-[4-(benzylcarbamoylamino)-2-[(2- hydroxy-l,l-dimethyl-ethyl)sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate.
Scheme 33:
B2Pin2, KOAc_
Pd(dppf)CI2, dioxane, 80°C
TBSCI
TEA, DCM
Figure imgf000149_0002
13 14
Figure imgf000150_0001
Preparation of compound 11.
Figure imgf000150_0002
10 11
To a mixture of 2-amino-2-methyl-propan-l-ol (3 g, 33.7 mmol, 5.1 eq.) and DMAP (80 mg, 654.8 umol, 0.98 eq.) in DCM (50 mL) was added 2-bromo-5-nitro- benzenesulfonyl chloride (2 g, 6.66 mmol, 1 eq.). The mixture was stirred at 20°C for 30 mins, then washed with IN HC1 (20 mL) and sat.aq.NaHC03 (20 mL). The organic layer was dried over Na2S04, filtered and concentrated to give 2-bromo-N-(2-hydroxy-l,l-dimethyl-ethyl)-5- nitro- benzenesulfonamide (1.7 g, 4.8 mmol, 72.3% yield) as a yellow gum. ESI [M+H] = 355.0/353.0
Preparation of compound 12.
Figure imgf000150_0003
11 12
A mixture of 2-bromo-N-(2-hydroxy-l,l-dimethyl-ethyl)-5-nitro- benzenesulfonamide (1.7 g, 4.8 mmol, 1 eq.), Fe (1.5 g, 26.9 mmol, 5.6 eq.) and NH4C1 (800 mg, 14.9 mmol, 3.1 eq.) in EtOH (15 mL)/H20 (7.5 mL)/THF (7.5 mL) was stirred at 80°C for 2 hrs. The reaction mixture was concentrated to remove EtOH, diluted with H20 (100 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by column chromatography (Si02, Petroleum ether: Ethyl acetate = 100: 1 to 1 : 1) to give 5-amino-2-bromo-N-(2-hydroxy-l,l-dimethyl- ethyl)benzenesulfonamide (1.1 g, 3.4 mmol, 70.7% yield) as a pale yellow solid. ESI [M+H] = 325.0/323.0
Preparation of compound 13.
Figure imgf000151_0001
12
A mixture of 5-amino-2-bromo-N-(2-hydroxy-l,l-dimethyl-ethyl) benzenesulfonamide (400 mg, 1.24 mmol, 1 eq.), 4,4,5,5-tetramethyl-2-(4,4,5,5- tetramethyl- l,3,2-dioxaborolan-2-yl)- 1,3,2-dioxaborolane (942.8 mg, 3.7 mmol, 3 eq.), Pd(dppf)Cl2 (90.6 mg, 123.8 umol, 0.1 eq.) and KOAc (364.4 mg, 3.7 mmol, 3 eq.) in dioxane (4 mL) was stirred at 80°C for 12 hrs under N2 atmosphere and then concentrated. The residue was purified by column chromatography (Si02, Petroleum ether: Ethyl acetate = 50: 1 to 3: 1) to give 5-amino-N-(2- hydroxy-1,1- dimethyl-ethyl)-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)benzenesulfonamide (201 mg, crude) as a white solid. 1H NMR (400MHz, METHANOL- d4) δ = 7.53 (d, J=8.0 Hz, 1H), 7.28 (d, J=2.4 Hz, 1H), 6.75 (dd, J=6.4 Hz, 1H), 3.35 (s, 2H), 1.36 (s, 12H), 1.10 (s, 6H).
Preparation of compound 14.
Figure imgf000151_0002
13 14
A mixture of 5-amino-N-(2-hydroxy-l,l-dimethyl-ethyl)-2-(4,4,5,5-tetramethyl -1,3,2- dioxaborolan-2-yl)benzenesulfonamide (140.7 mg, 380.1 umol, 1.2 eq.), trans-isopropyl N- [4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (110 mg, 316.7 umol, 1 eq.), Na2C03 (100.7 mg, 950.3 umol, 3 eq.), KF (27.6 mg, 475.1 umol, 1.5 eq.) and Pd(PPh3)4 (36.6 mg, 31.7 umol, 0.1 eq.) in toluene(l mL)/EtOH (1 mL)/H20 (0.3 mL) was stirred at 80°C for 12 hrs under N2 atmosphere. The reaction mixture was concentrated, diluted with H20 (10 mL) and extracted with EtOAc (10 mL x 2). The combined organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by prep-TLC (Si02, Ethyl acetate) to give trans-isopropyl N-[4-[5-[4-amino-2-[(2-hydroxy-l,l-dimethyl- ethyl)sulfamoyl] phenyl]thiazol-2-yl]cyclohexyl]carbamate (117 mg, crude) as a yellow solid. 1H NMR (400MHz, METHANOL-d4) δ = 7.63 (d, J=7.0 Hz, 1H), 7.44 (d, J=2.2 Hz, 1H), 7.15 (d, J=8.3 Hz, 1H), 6.83 (dd, J=2.4, 8.1 Hz, 1H), 4.85 - 4.78 (m, 1H), 3.50 - 3.39 (m, 1H), 3.25 (s, 2H), 3.03 - 2.91 (m, 1H), 2.21 (br t, J=6.1 Hz, 2H), 2.12 - 2.00 (m, 2H), 1.75 - 1.60 (m, 2H), 1.45 - 1.33 (m, 2H), 1.28 - 1.19 (s, 6H), 1.03 (s, 6H)
Preparation of compound 15.
Figure imgf000152_0001
To a solution of trans-isopropyl N-[4-[5-[4-amino-2-[(2-hydroxy-l,l-dimethyl-ethyl) sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate (90 mg, 176.3 umol, 1 eq.) in DCM (1 mL) were added TEA (53.5 mg, 528.7 umol, 3 eq.), DMAP (2.2 mg, 17.6 umol, 0.1 eq.) and TBSC1 (66.4 mg, 440.6 umol, 2.5 eq.). The mixture was stirred at 30°C for 12 hrs and then concentrated. The residue was purified by prep-TLC (Si02, Petroleum ether: Ethyl acetate = 2: 1) to give trans-isopropyl N-[4-[5-[4-amino-2-[[2-[tert-butyl(dimethyl)silyl]oxy-l,l- dimethyl-ethyl]sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate (120 mg, crude) as a pale yellow solid. ESI [M+H] = 625.2
Preparation of compound 16.
Figure imgf000152_0002
To a solution of trans-isopropyl N-[4-[5-[4-amino-2-[[2-[tert-butyl(dimethyl)silyl] oxy-1,1- dimethyl-ethyl]sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate (90 mg, 144 umol, 1 eq.) in DCM (1 mL) were added DMAP (1.8 mg, 14.4 umol, 0.1 eq.), pyridine (34.2 mg, 432 umol, 3 eq.) and (4-nitrophenyl) carbonochloridate (43.5 mg, 216 umol, 1.5 eq.). The mixture was stirred at 25°C for 0.5 hr and used into the next step directly without further purification. ESI [M+H] = 790.3 Preparation of compound 17.
Figure imgf000153_0001
16 17
To a solution of phenylmethanamine (36 mg, 336 umol, 3 eq.) in DCM (1 mL) was added a solution of trans-(4-nitrophenyl) N-[3-[[2-[tert-butyl(dimethyl)silyl] oxy-l,l-dimethyl- ethyl]sulfamoyl]-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5- yljpheny I] carbamate (88.5 mg, 112 umol, 1 eq.) in DCM (1 mL) and the mixture was stirred at 25 °C for 0.5 hr. The reaction mixture was diluted with H20 (10 mL) and extracted with EtOAc (10 mL x 2). The combined organic layers were dried over Na2S04, filtered and concentrated. The residue was purified by prep-TLC (Si02, Petroleum ether: Ethyl acetate =
1 : 1) to give trans-isopropyl N-[4-[5-[4-(benzylcarbamoylamino)-2-[[2-[tert- butyl(dimethyl)silyl]oxy-l,l-dimethyl-ethyl]sulfamoyl]phenyl]thiazol-2- yl]cyclohexyl]carbamate (93 mg, crude) as a yellow gum. ESI [M+H] = 758.4
Preparation of Ex. 74.
Figure imgf000153_0002
17 Ex. 74
A mixture of trans-isopropyl N-[4-[5-[4-(benzylcarbamoylamino)-2-[[2-[tert- butyl(dimethyl)silyl]oxy-l,l-dimethyl-ethyl]sulfamoyl]phenyl]thiazol-2- yl]cyclohexyl]carbamate (84.9 mg, 112 umol, 1 eq.) in AcOH (0.5 mL)/THF (0.5 mL)/H20 (0.5 mL) was stirred at 80°C for 0.5 hr. Then the mixture was concentrated and the residue was purified by prep-HPLC (TFA condition) to give trans-isopropyl N-[4-[5-[4- (benzylcarbamoylamino )-2-[ ( 2-hydroxy-l,l-dimethyl-ethyl)sulfamoyl]phenyl]thiazol-2- yl]cyclohexyl]carbamate (5.86 mg, 9.10 umol, 8.13% yield, 100% purity) as a yellow solid. 1H NMR (400MHz, METHANOL-d4) δ = 8.25 (d, J=2.6 Hz, 1H), 7.81 - 7.66 (m, 2H), 7.40 - 7.30 (m, 5H), 7.28 - 7.22 (m, 1H), 4.82 (td, J=6.0, 12.5 Hz, 1H), 4.41 (s, 2H), 3.45 (br t, J=11.6 Hz, 1H), 3.28 (s, 2H), 3.00 (br t, J=11.8 Hz, 1H), 2.23 (br d, J=13.2 Hz, 2H), 2.11 - 2.00 (m, 2H), 1.77 - 1.60 (m, 2H), 1.48 - 1.32 (m, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.05 (s, 6H). ESI [M+H] = 644.3
Example 75 Compound Primary Screening
1. BACKGROUND
Primary screening was a phenotypic screen that utilized the synthetic lethal interaction between AID and RAD51 to identify compounds that were both potent and on target. AID expressing cells are dependent upon RAD51 for survival; inhibiting RAD51 in AID positive cells results in a cytotoxic effect. Based on such an effect, compounds that were potent in AID positive cells and were signficiantly less potent in AID negative cells were identified.
2. MATERIALS AND SUPPLIES
Plastic ware and consumables needed for this experiment include: Cell Culture media; Evaporation Buffer media; 100% DMSO; 96 well U-bottom sterile culture plates; 250mL bottle; 1.5mL Opaque amber epi tubes; Epi Tube rack; 300mL reservoirs; 25mL reservoir; 25mL serological pipette tips; 5mL serological pipette tips P1000 Pipette Tips; and P200 Pipette Tips.
Equipment needed for this experiment include: Viaflo 384 liquid handler; Eppendorf serological pipette; Eppendorf PI 000 Pipette; and Eppendorf P200 Pipette
Daudi Cell Culture and WI-38 Cell Cultures were also needed for this experiment. Lastly, compounds (e.g. , the compounds of this invention) to be tested are needed.
3. PROCEDURE
All steps were performed in a sterile environment inside the Biosafety cabinet.
The first step was to set up a cell killing assay in the Daudi cell line (AID positive). A 96 well u-bottom plate was prepared by writing the experiment number, plate number, date and initials in the top right corner of the plate lid. With a sterile 300ml reservoir, and 25ml serological pipette, evaporation buffer media was pipetted into reservoir in 25ml increments. Using the liquid handler, 150ul of evaporation buffer media was pipetted from reservoir into rows A and H, and Columns 1 and 12 of the 96 well u-bottom plate. Cell cultures were counted to obtain the density of cells per ml, and the culture viability. The cell density information was used to obtain 1,000,000 cells from culture using a 5mL serological pipette into an epi tube. The cell density information from the culture was used to calculate the number of cells and volume of media needed for the assay to seed 1250 cells in 130ul of media per available culture well in the 96 well u-bottom plate. Rows B through F were used for cells (50 wells in total), with row G left for an empty media control. The calculation was overestimated by lOmL to account for the dead volume in the 300ml reservoir. Once the media volume was calculated, the appropriate volume of media was pipetted in 25mL increments into the 250mL bottle using a 25mL serological pipette. The 250ml bottle was capped tightly, and placed into a 37°C water bath for 2 minutes. While the culture media was warming, lOmL of fresh media was pipetted from the 500mL culture media bottle into a sterile 25mL reservoir. Using the Eppendorf multichannel pipette, 130ul of media was piptted from the 25mL reservoir into row G of the 96 well u-bottom plate. Once the 250mL bottle of media was warmed, the volume of culture needed was pipetted into the bottle, and mixed gently with a 25mL serological pipette as to not create bubbles, and then the contents of the bottle were pipetted into a new 300mL reservoir. Using the liquid handler, 130ul of culture was pipetted from the 300mL reservoir into rows B through F of the 96 well u-bottom plate. Once the culture was added, the plate was placed into a 37°C incubator until the compound master plate was prepared for use.
Two 96 well u-bottom plates were prepared by writing the master plate name in the upper right corner of the plate lid. Labeling one DMSO master and the other Media Master. The compounds of interest were obtained from the laboratory freezer, and placed into a 25 well storage box with a lid, and set the box aside. The compounds were vortexed after thawing but before use. Using an automatic multichannel pipette, 20ul of 100% DMSO was pipetted into wells B3-B 11 through G3-G11 of the DMSO master plate. For each compound on the master plate, 50ul of the compound were pipetted in the appropriate well of row 2 (reference plate map to determine appropriate well). A serial dilution was prepared beginning by aspirating 20ul from row 2 and mixing with row 3, repeating until row 11 was reached. Using the liquid handler, 194ul of Daudi media was dispensed into wells B2-B 11 through G2-G11 of the Media master plate. Using the liquid handler, 6ul from the DMSO master plate was aspirated and dispensed into the media master plate, mixing lOOul twice.
Compounds from master plate were then added to the culture plate. The culture plates were removed from the incubator, and set inside the biosafety cabinet. Using a liquid handler, 20ul from wells B2 to B 11 through G2 to Gl 1 of master plate were aspirated, and dispensed into wells B2 to B 11 through G2 to Gl 1 of culture plate. This set was continued with each culture plate. Once the culture plates acquired their 20ul of compound dilutions, they were placed back into the incubator, until their reads on Day 7 of experiment. Cell death was measured on Day 7 of the experiment using Cell-Titer Glo and a Promega Plate reader. Percent cell death and EC50 values were calculated by comparing the cell viability of the compound treated wells to the non- treated wells. Normalized RLU values were obtained by subtracting the media well values from each of the wells in the same column, and then dividing that value by the DMSO treated cells values. The percent kill was then calculated by subtracting the normalized RLU value from 1 and multiplying by 100. The average normalized percent kill value and standard error of the mean was then calculated. The kill values were then inputted into Prism with the corresponding standard errors. In Prism a nonlinear regression line was plotted with the data points using a semi- log scale, and the EC50 value was calculated. For compounds that showed good potency in the Daudi cell line, the assay was repeated using WI-38 cells (AID negative).
Screening Data
Table 1:
Figure imgf000156_0001
Figure imgf000157_0001
Figure imgf000158_0001
Figure imgf000159_0001

Figure imgf000160_0001
Figure imgf000161_0001
Figure imgf000162_0001
Figure imgf000163_0001
Figure imgf000164_0001
Figure imgf000165_0001
Figure imgf000166_0001
Example 76. Bi-directional Caco-2 permeability
Bi-directional Caco-2 permeability was assayed. Caco-2 cells were seeded onto permeable polycarbonate supports and allowed to differentiate for about 3 weeks prior to being used in the assays. The cells were then exposed to the compounds from either the apical or basolateral sides and incubated at 37C for up to 90 minutes under light agitation. Compound transport was then measured using LC/MS/MS analysis at 30, 60, and 90 minutes.
Table 2
Figure imgf000166_0002
1.9 3.3 1.7 66.9 81
Ex. 28
10.9 24.1 2.2 93.9 90.9
Ex. 29
11.5 15.7 1.4 79.8 115.6
Ex. 30
12.8 13.6 1.1 70.5 92.3
Ex. 31
0.4 45.2 103.9 98.2 96.8
Ex. 34
0.4 41 99.1 98.8 107.5
Ex. 35
17.9 22.9 1.3 73.3 82.3
Ex. 38
3.2 4.2 1.3 36.2 69.4
Ex. 44
7.6 10.8 1.4 73.6 84.4
Ex. 45
8.7 12.4 1.4 65.3 80.7
Ex. 47
23.1 16 0.7 80.1 90.3
Ex. 48
2 29.1 14.6 87.7 98.1
Ex. 50
9.9 10.5 1.1 65 85.7
Ex. 51
7.4 10.9 1.5 64.1 91.7
Ex. 52
9.3 9.7 1 63.1 90.9
Ex. 53
6.5 7 1.1 65.2 83.8
Ex. 55
1.3 3.5 2.7 61.9 86.8
Ex. 57
5.1 3.4 0.7 52.3 83.5
Ex. 58
3.1 15 4.8 61.5 91.5
Ex. 59
3.1 9.6 3.1 55.6 85.5
Ex. 59A
4.6 9.7 2.1 56.2 82
Ex. 59B
15.6 13.3 0.9 59.8 90.8
Ex. 60
12 11.2 0.9 55.3 86
Ex. 60A
11.6 13.5 1.2 51.2 84.3
Ex. 60B
10.5 26.7 2.5 83.6 90
Ex. 63
2.2 27.4 12.5 79.7 95.9
Ex. 64 2.9 3.9 1.4 64.9 81.2
Ex. 66A
10.5 6.9 0.7 50.7 78.6
Ex. 67A
Example 77. Human Liver Microsome Stability
The stability of the claimed compounds was determined in the presences of human liver microsomes. The compounds were incubated with the microsomes at 37 °C for 45 minutes. Samples were analyzed using LC-MS/MS. Data analysis included half- life, clearance rate, and the percentage of hepatic blood flow (%QH) for each of the compounds in the different species. Below are liver microsome assat data of representative compounds, which show that the claimed compounds have superior metabolic stability.
Table 3
Figure imgf000168_0001
Example 78. Cell Line Screen
The activity of the claimed compounds was measured in a variety of cell lines with different expression levels of activation induced cytidine deaminase (AICDA). The potency assay was repeated in all of the listed cell lines and the EC50 values recorded.
Table 4
Figure imgf000168_0002
(Head and Neck)
SNU-5
Low n.d. n.d. 2941 3845
(Gastric)
TOV-1120D
Negative 9172 n.d. 2377 4924
(Ovary)
OV56
Low 9086 n.d. 5944 7228
(Ovary)
ARPE19/HPV16
(HPV Immortalized Negative >10000 >10000 >10000 >10000 RPE)
WI-38 (Normal
Human Lung Negative >10000 >10000 >10000 >10000 Fibroblast)
n.d. not determined
Example 79. Pharmacokinetic (PK)
PK studies in mice were used to determine the fate of the compounds in a whole organism. Rats were treated with the compounds either orally of via IV at the indicated doses and followed for up to 24 hours. Plasma samples were taken at different time points and analyzed by LC-MS.
Table 5
Figure imgf000169_0001

Claims

What is claimed is:
1. A compound represented by the following structural formula:
Figure imgf000170_0001
or a pharmaceutically acceptable salt thereof, wherein:
the thiazole ring is optionally substituted with -F or -CI;
Cy is -(C3-C7)cycloalkyl, bridged (C6-C12) cycloalkyl, or a 4-10 membered heterocyclic ring, each of which is optionally substituted with one or more groups selected from the group consisting of halogen, -OH, (Ci-C4)alkyl, and (Ci-C4)alkoxy; when X5 is connected with a nitrogen ring atom of Cy, X5 is absent;
when X5 is connected with a carbon ring atom of Cy, X5 is NR or O;
X6 is NRa or O;
R1 is (Ci-C5)alkyl optionally substituted with -OH;
R3 is (Ci-C5)alkyl, -CH2-phenyl, -(C3-C7)cycloalkyl, -CH2- (C3-C7)cycloalkyl,
-CH2-monocyclic 3-7 membered heterocyclic ring, or monocyclic 3-7 membered heterocyclic ring, wherein the (Ci-C5)alkyl, -(C3-C7)cycloalkyl, phenyl or monocyclic 3-7 membered heterocyclic ring represented by R or in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen, -OH, (Ci-C4)alkyl, halomethyl, halomethoxy, -CN, and (Ci-C4)alkoxy;
R2 is -NRaC(0)0(Ci-C4)alkyl; -NRaC(0)NRa(Ci-C4)alkyl; -NRaC(0)0(C2- C4)alkenyl; -NRaC(0)NRa(C2-C4)alkenyl; -NRaC(0)0-(C3-C6)cycloalkyl;
-NRaC(0)NRa-(C3-C7)cycloalkyl; -NRaC(0)0-phenyl; -NRaC(0)NRa-phenyl;
-NR C(0)0-monocyclic 3-7 membered heterocyclic ring; -NR C(0)NR -monocyclic 3-7 membered heterocyclic ring; -NR C(0)0-monocyclic 5-6 membered
heteroaromatic ring; -NR C(0)NR -monocyclic 5-6 membered heteroaromatic ring; wherein the (Ci-C4)alkyl and the (C2-C4)alkenyl in the group represented by
R are each optionally and independently substituted with one or more groups selected from the group consisting of halogen, N3, -OR , -NR R , -(C3-C6)cycloalkyl, phenyl, a monocyclic 3-7 membered heterocyclic ring, and a monocyclic 5-6 membered heteroaromatic ring;
wherein the (C3-C7)cycloalkyl in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen, -CH3, =0, -ORa and -NRaRa;
wherein the phenyl in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen, -CH3, halo methyl, halomethoxy, -CN, -OR , and -N3;
wherein the heterocyclic ring in the group represented by R is optionally substituted with one or more groups selected from the group consisting of =0, halogen, -OR , -CH3, halomethyl, and halomethoxy;
wherein the heteroaromatic ring in the group represented by R is optionally substituted with one or more groups selected from the group consisting of halogen, -CN, -CH3, halomethyl, halomethoxy, -ORa and -NRaRa; and
each R is independently -H or -CH3.
2. A compound represented by the following structural formula:
Figure imgf000171_0001
or a pharmaceutically acceptable salt thereof, wherein:
the thiazole ring is optionally substituted with -F or -CI;
Cy is cyclohexyl or a 6-membered monocyclic heterocyclic ring;
X5 and X6 are each independently NR or O;
R1 is (Ci-C5)alkyl;
R is (Ci-C5)alkyl or monocyclic 3-7-membered heterocyclic ring;
R2 is -NRaC(0)0(Ci-C4)alkyl; -NRaC(0)NRa(Ci-C4)alkyl; -NRaC(0)0(C2- C4)alkenyl; -NRaC(0)NRa(C2-C4)alkenyl; -NRaC(0)-0(C3-C6)cycloalkyl; -NRaC(0)NRa-(C3-C6)cycloalkyl; -NRaC(0)0-phenyl; -NRaC(0)NRa-phenyl;
-NR C(0)0-monocyclic 3-7 membered heterocyclic ring; -NR C(0)NR -monocyclic 3-7 membered heterocyclic ring; -NR C(0)0-monocyclic 5-6 membered
heteroaromatic ring; -NR C(0)NR -monocyclic 5-6 membered heteroaromatic ring; wherein the (Ci-C4)alkyl and the (C2-C4)alkenyl in the group represented by
R are each optionally and independently substituted with one or more halogen, N3, -OR , -NR R , -(C3-C6)cycloalkyl, phenyl, monocyclic 3-7-membered heterocyclic ring, or monocyclic 5-6-membered heteroaromatic ring;
wherein the -(C3-C6)cycloalkyl in the group represented by R is optionally substituted with one or more halogen, -CH3, -OR or -NR R ;
wherein the phenyl in the group represented by R is optionally substituted with one or more halogen, -CH3, halomethyl, halomethoxy, -OR , or -N3;
wherein the heterocyclic ring in the group represented by R is optionally substituted with one or more =0, halogen, -CH3, halomethyl, or halomethoxy;
wherein the heteroaromatic ring in the group represented by R is optionally substituted with one or more halogen, -CH3, halomethyl, halomethoxy, -OR or -NRaRa; and
each R is independently -H or -CH3.
3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein Cy is
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl; azetidinyl, azepanyl, diazaspiro[4.4]nonyl, diazaspiro[3.5]nonyl, diazepanyl, dihydroimidazole, dihydrofuranyl, dihydropyranyl, dihydropyridinyl,
dihydropyrimidinyl, dihydrothienyl, dihydrothiophenyl, dihydrothiopyranyl, hexahydropyridazinyl, hexahydropyrimidinyl, hydantoinyl, indolinyl, isoindolinyl, morpholinyl, oxiranyl, oxetanyl, piperidinyl, piperazinyl, pyrrolidinonyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydroimidazole, tetrahydroindolyl, tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridinyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, thio morpholinyl, tropanyl, valerolactamyl;
bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl, bicyclo[4.3.1]decyl, bicyclo[3.3.1]nonyl, bornyl, bornenyl, norbornyl, norbornenyl, 6,6-dimethylbicyclo [3.1.1]heptyl, tricyclo butyl, adamantly; azanorbornyl, quinuclidinyl, isoquinuclidinyl, tropanyl,
azabicyclo [2.2. l]heptanyl, 2-azabicyclo[3.2. l]octanyl, azabicyclo [3.2. l]octanyl, azabicyclo[3.2.2]nonanyl, azabicyclo[3.3.0]nonanyl, azabicyclo [3.3.1] no nanyl, diazabicyclo[2.2. l]heptanyl, diazabicyclo[3.2. l]octanyl, octahydropyrrolo[3,4- b]pyrrolyl, octahydropyrrolo[3,4-c]pyrrolyl.
4. The compound of any one of claims 1-3, or a pharmaceutically acceptable salt
thereof, wherein Cy is cyclohexyl, morpholinyl, thiomorpholinyl, piperidinyl, piperazinyl, hexahydropyridazinyl, hexahydropyrimidinyl, valerolactamyl, dihydropyranyl, dihydropyridinyl, dihydropyrimidinyl, dihydrothiopyranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydropyrimidinyl, or
tetrahydrothiopyranyl.
5. The compound of any one of claims 1-4, represented by the following structural formula:
Figure imgf000173_0001
or a pharmaceutically acceptable salt thereof, wherein:
X7 is NH or O;
R4 is (Ci-C4)alkyl, (C3-C6)cycloalkyl, or a monocyclic 3-7 membered heterocyclic ring;
wherein the (Ci-C4)alkyl represented by R4 is optionally substituted with one or more groups selected from the group consisting of halogen, N3, -OR , -NR R , -(C3-C6)cycloalkyl, phenyl, a monocyclic 3-7 membered heterocyclic ring, and a monocyclic 5-6 membered heteroaromatic ring,
wherein the (C3-C6)cycloalkyl or the monocyclic 3-7 membered heterocyclic ring represented by R4, the (C3-C6)cycloalkyl or the monocyclic 3-7 membered heterocyclic ring in the group represented by R4 is optionally substituted with one or more groups selected from the group consisting of halogen, -OR , =0, and -CH3, wherein the phenyl in the group represented by R4 is optionally substituted with one or more groups selected from the group consisting of halogen, -CH3, halomethyl, halomethoxy, -OR , and -N3;
wherein the heteroaromatic ring in the group represented by R4 is optionally substituted with one or more groups selected from the group consisting of halogen and -CH3.
The compound of claim 5 or a pharmaceutically acceptable salt thereof, wherein X7 is NH or O;
R3 is (Ci-C5)alkyl; and
R4 is (Ci-C4)alkyl wherein the (Ci-C4)alkyl represented by R4 is optionally substituted with one or more halogen, -OR , -NR R , -(C3-C6)cycloalkyl, phenyl (optionally substituted by one or more halogen, -CH3, halomethyl, halomethoxy, OR' or N3), monocyclic 3-7-membered heterocyclic ring (optionally substituted by =0, halogen or -CH3), or monocyclic 5-6-membered heteroaromatic ring (optionally substituted by halogen or -CH3).
The compound any one of claims 1-6, represented by the following structural formula:
Figure imgf000174_0001
or a pharmaceutically acceptable salt thereof.
8. The compound any one of claims 1-6, represented by the following structural
formula:
Figure imgf000174_0002
or a pharmaceutically acceptable salt thereof.
9. The compound any one of claims 1-6, represented by the following structural formula:
Figure imgf000175_0001
or a pharmaceutically acceptable salt thereof.
The compound any one of claims 1-6, represented by the following structural formula:
Figure imgf000175_0002
or a pharmaceutically acceptable salt thereof.
11. The compound any one of claims 1-6, represented by the following structural formula:
Figure imgf000175_0003
or a pharmaceutically acceptable salt thereof.
The compound any one of claims 1-6, represented by the following structural formula:
Figure imgf000175_0004
or a pharmaceutically acceptable salt thereof.
13. The compound any one of claims 1-6, or a pharmaceutically acceptable salt thereof, wherein Cy is azetidinyl or pyrrolidinyl, and the nitrogen ring atom is connected with the thiazole ring.
14. The compound any one of claims 1-6, or a pharmaceutically acceptable salt thereof, wherein Cy is l,7-diazaspiro[4.4]nonyl, 2,7-diazaspiro[4.4]nonyl,
2.7- diazaspiro[3.5]nonyl, 1,4-diazepanyl, 2,5-diazabicyclo[2.2. l]heptanyl,
3.8- diazabicyclo[3.2. l]octanyl, octahydropyrrolo[3,4-b]pyrrolyl, or
octahydropyrrolo[3,4-c]pyrrolyl, and the two nitrogen ring atoms are connected with the thiazole ring and the -X5C(0)X6R3 moiety, respectively.
15. The compound of any one of claims 5 and 7-14, or a pharmaceutically acceptable salt thereof, wherein R4 is -(Ci-C3)alkyl, (C3-C6)cycloalkyl, or a monocyclic 3-7 membered heterocyclic ring, wherein the -(Ci-C3)alkyl is optionally substituted with
(i) phenyl optionally substituted by one or more halogen or -CI¾; (ii) a monocyclic 5-6 membered heteroaromatic ring optionally substituted by one or more halogen or -CH3; or (iii) a monocyclic 3-7 membered heterocyclic ring optionally substituted by one or more groups selected from the group consisting of halogen and -CH3.
16. The compound of any one of claims 5 and 7-14, or a pharmaceutically acceptable salt thereof, wherein R4 is -(Ci-C3)alkyl, -CHRa-phenyl, -CHRa-5-6 membered heteraromatic ring, or -CHR -3-7 membered monocyclic heterocyclic ring, wherein the phenyl, 5-6 membered heteraromatic ring or 3-7 membered monocyclic heterocyclic ring in the group represented by R4 is optionally substituted one or more groups selected from the group consisting of halogen and -CH3.
17. The compound of any one of claims 5-14, or a pharmaceutically acceptable salt
thereof, wherein R4 is -(Ci-C3)alkyl, optionally substituted with (i) phenyl optionally substituted by one or more halogen, -CH3, halomethyl, halomethoxy, OR , or N3;
(ii) a monocyclic 5-6-membered heteroaromatic ring optionally substituted by one or more halogen or -CH3; or (iii) a monocyclic 3-7-membered heterocyclic ring optionally substituted by one or more =0 or -CH3.
18. The compound of claim 17, or a pharmaceutically acceptable salt thereof, wherein R4 is (i) -(Ci-C3)alkyl; (ii) -CH2-phenyl optionally substituted by halogen, -CH3, halo methyl, halomethoxy, OR , or N3; (iii) -CH(CH3)-phenyl optionally substituted by halogen, -CH3, halomethyl, halomethoxy, OR , or N3; (iv) -CH2-5-6-membered heteraromatic ring optionally substituted by halogen or -CH3; or (v) -CH2-3-7- membered monocyclic heterocyclic ring optionally substituted by =0 or -CH3.
19. The compound of any one of claims 1, 3-5, and 7-18, or a pharmaceutically
acceptable salt thereof, wherein R is (Q-G alkyl, -(C4-C6)cycloalkyl, -CH2-phenyl, -CH2-monocyclic 4-6 membered heterocyclic ring, or monocyclic 4-6 membered heterocyclic ring, wherein the phenyl or monocyclic 4-6 membered heterocyclic ring represented by R 3 or in the group represented by R 3 is optionally substituted with one or more groups selected from the group consisting of halogen, -OR , and -CH3.
20. The compound of any one of claims 1-7 and 15-19, represented by the following structural formula:
Figure imgf000177_0001
or a pharmaceutically acceptable salt thereof.
The compound of claim 20, represented by the following structural formula:
Figure imgf000177_0002
or a pharmaceutically acceptable salt thereof. The compound of any one of claims 1-6, 8, and 15-19, represented by the following structural formula:
Figure imgf000178_0001
or a pharmaceutically acceptable salt thereof.
The compound of claim 22, represented by the following structural formula:
Figure imgf000178_0002
or a pharmaceutically acceptable salt thereof.
The compound of any one of claims 1-6, 10, and 15-19, represented by the following structural formula:
Figure imgf000178_0003
or a pharmaceutically acceptable salt thereof.
25. The compound of any one of claims 1-24, or a pharmaceutically acceptable salt
thereof, wherein R is isopropyl, ieri-butyl, cyclobutyl, cyclopentyl, benzyl, oxetanyl,
tetrahydro-2H-pyranyl,
Figure imgf000178_0004
26. The compound of any one of claims 1-25, or a pharmaceutically acceptable salt thereof, wherein R1 is tert-butyl.
27. The compound of any one of claims 1-26, or a pharmaceutically acceptable salt thereof, wherein R is isopropyl or oxetanyl.
28. The compound of claim 27, or a pharmaceutically acceptable salt thereof, wherein R is isopropyl.
29. The com ound of any one of claims 5-28, or a pharmaceutically acceptable salt
Figure imgf000179_0001
31. The compound of claim 29, or a pharmaceutically acceptable salt thereof, wherein R4
Figure imgf000179_0002
The compound of claim 29, or a pharmaceutically acceptable salt thereof, wherein R4
Figure imgf000180_0001
The compound of claim 29, or a pharmaceutically acceptable salt thereof, wherein R4
Figure imgf000180_0002
The compound of claim 29, or a pharmaceutically acceptable salt thereof, wherein R4
Figure imgf000180_0003
35. A pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and a compound of any one of claims 1-34 or a pharmaceutically acceptable salt thereof.
36. A method of treating cancer, autoimmune disease, immune deficiency, or
neurodegenerative disease, the method comprising administering to a subject in need thereof an effective amount of a compound of any one of claims 1-34 or a
pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 35.
37. The method of claim 36, wherein the method is a method of treating cancer and the cancer is selected from the group consisting of lymphoma, leukemia, and a plasma cell neoplasm.
38. The method of claim 37, wherein the cancer is lymphoma selected from the group consisting of Non-Hodgkin's lymphoma; Burkitt's lymphoma; small lymphocytic lymphoma; lymphoplasmacytic lymphoma; MALT lymphoma; follicular lymphoma; diffuse large B-cell lymphoma; and T-cell lymphoma.
39. The method of claim 37, wherein the cancer is leukemia selected from the group
consisting of acute lymphoblastic leukemia (ALL); Burkitt's leukemia; B- cellieukemia; B-cell acute lymphoblastic leukemia; chronic lymphocytic leukemia (CLL); acute myelogenous leukemia (AML); chronic myelogenous leukemia (CML); and T-cell acute lymphoblastic leukemia (T-ALL).
40. The method of claim 37, wherein the cancer is plasma cell neoplasm selected from the group consisting of multiple myeloma; plasma cell myeloma; plasma cell leukemia; and plasmacytoma.
41. The method of claim 36, wherein the method is a method of treating cancer and the cancer is selected from the group consisting of carcinoma and sarcoma.
42. The method of claim 41, wherein the cancer is carcinoma selected from the group consisting of colon cancer; liver cancer; gastric cancer; intestinal cancer; esophageal cancer; breast cancer; ovarian cancer; head and neck cancer; lung cancer; and thyroid cancer
43. The method of claim 36, wherein the method is a method of treating cancer and the cancer is selected from the group consisting of colon cancer, endometrial cancer, stomach cancer, pancreatic cancer, kidney/ureter tract cancer, hepatobiliary tract cancer, gastric tract cancer, prostate cancer, ovarian cancer, gallbladder duct cancer, brain cancer, small intestine cancer, breast cancer, and skin cancer.
44. The method of any one of claims 36-43, wherein the method is a method of treating cancer and the cancer is characterized by mutations in mutS homologue 6 (MSH6).
45. The method of claim 36, wherein the method is a method of treating autoimmune disease selected from the group consisting of lupus erythematosus; Wiskott-Aldrich syndrome; autoimmune lymphoproliferative syndrome; myasthenia gravis;
rheumatoid arthritis (RA); lupus nephritis; systemic lupus erythematosis; discoid lupus; subacute cutaneous lupus erythematosus; cutaneous lupus erythematosus including chilblain lupus erythematosus; chronic arthritis; Sjogren's syndrome;
inflammatory chronic rhinosinusitis; colitis; celiac disease; inflammatory bowel disease; Barrett's esophagus; inflammatory gastritis; autoimmune nephritis;
autoimmune vasculitis; autoimmune hepatitis; autoimmune carditis; autoimmune encephalitis; autoimmune diabetes; autoimmune diabetes nephritis; psoriasis; Graft- versus-host disease (GvHD); and autoimmune mediated hematological disease.
46. The method of claim 36, wherein the method is a method of treating immune
deficiency selected from the group consisting of Autoimmune Lymphoproliferative Syndrome (ALPS), Autoimmune polyglandular syndrome type 1 (APS-1), BENTA Disease, Caspase Eight Deficiency State (CEDS), Chronic Granulomatous Disease (CGD), Common Variable Immunodeficiency (CVID), Congenital Neutropenia Syndromes, CTLA4 Deficiency, DOCK8 Deficiency, GATA2 Deficiency,
Glycosylation Disorders With Immunodeficiency, hyper-immunoglobulin E syndrome (HIES), Hyper-Immunoglobulin M (Hyper-IgM) Syndromes, Leukocyte adhesion deficiency (LAD), LRBA deficiency, PI3 Kinase disease, PLCG2-associated antibody deficiency and immune dysregulation (PLAID), severe combined immunodeficiency (SCID), STAT3 gain-of-function disease, Warts,
Hypogammaglobulinemia, Infections, and Myelokathexis Syndrome (WHIMS), X- Linked Agammaglobulinemia (XLA), X-Linked Lymphoproliferative Disease (XLP), and XMEN Disease.
47. The method of claim 36, wherein the method is a method of treating a
neurodegenerative disorder selected from the group consisting of multiple sclerosis, Parkinson's disease (PD), Alzheimer's disease (AD), Dentatorubropallidoluysian atrophy (DRPLA), Huntington's Disease (HD), Spinocerebellar ataxia Type 1 (SCA1), Spinocerebellar ataxia Type 2 (SCA2), Spinocerebellar ataxia Type 3 (SCA3), Spinocerebellar ataxia 6 (SCA6), Spinocerebellar ataxia Type 7 (SCA7), Spinocerebellar ataxia Type 8 (SCA8), Spinocerebellar ataxia Type 12 (SCA12), Spinocerebellar ataxia Type 17 (SCA17), Spinobulbar Muscular Ataxia/Kennedy Disease (SBMA), Fargile X syndrome (FRAXA), Fragile XE mental retardation (FRAXE), and Myotonic dystrophy (DM).
48. The method of any one of claims 36-47, further comprising the step of coadministering to the subject an effective amount of a DNA repair inhibitor, a DNA damage response (DDR) inhibitor, a DNA damaging agent or an immunomodulatory agent.
49. The method of claim 48, wherein the DNA damaging agent is selected from the group consisting of: exposure to a DNA damaging chemical; exposure to a
chemotherapeutic agent; exposure to a radiochemotherapy, and exposure to ionizing or ultraviolet radiation.
50. The method of any one of claims 36-49, wherein the subject is determined to have an increased level and/or activity of a DNA damage process or DNA editing enzyme.
51. The method of claim 48, wherein the immunomodulatory agent is selected from the group consisting of immune checkpoint modulators, Toll-like receptor (TLR) agonists, cell-based therapies, cytokines and cancer vaccines.
52. The method of claim 50, wherein the DNA editing enzyme is selected from the group consisting of activation induced cytidine deaminase (AID or AICDA), APOBEC2, APOBEC3A, APOBEC3C, APOBEC3D, APOBEC3F, APOBEC3G, APOBEC3H, APOBEC4, a Type 1 Topoisomerase, a Type 2 Topoisomerase, Recombination Activating Gene 1 (RAG 1), and Recombination Activating Gene 2 (RAG2).
53. The method of any one of claims 36-52, wherein blood cells obtained from the
subject have been determined to have a detectable level of activation-induced cytidine deaminase (AID).
54. The method of any one of claims 36-52, wherein B cells obtained from the subject have been determined to have a detectable level of activation-induced cytidine deaminase (AID).
55. The method of claim 53 or 54, wherein the detectable level of activation-induced cytidine deaminase (AID) is statistically significantly higher than the level of AID expressed in unactivated B-cells or normal non-immune cells from a healthy subject.
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020257752A1 (en) * 2019-06-21 2020-12-24 Cyteir Therapeutics, Inc. Methods of using rad51 inhibitors for treatment of pancreatic cancer
WO2021164746A1 (en) 2020-02-19 2021-08-26 江苏先声药业有限公司 Substituted aryl compound
US20210275503A1 (en) * 2020-03-03 2021-09-09 Cyteir Therapeutics, Inc. Targeting homologous recombination: a new sythetic lethal therapeutic paradigm
EP3938358A4 (en) * 2019-03-12 2022-12-21 Cyteir Therapeutics, Inc. Rad51 inhibitors

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3652159B1 (en) 2017-07-11 2023-01-18 Cyteir Therapeutics, Inc. Substituted thiazole derivatives as rad51 inhibitors for the treatment of cancer diseases, autoimmune and neurodegenerative conditions
EP4112616A1 (en) * 2017-09-11 2023-01-04 Cyteir Therapeutics, Inc. Rad51 inhibitors
CN114173781A (en) 2019-03-25 2022-03-11 赛泰尔治疗公司 Combination of RAD51 and PARP inhibitors
JP2022151436A (en) * 2021-03-26 2022-10-07 均 石井 Agents for treating meningitidis and encephalitis
WO2023078271A1 (en) * 2021-11-02 2023-05-11 上海旭成医药科技有限公司 Aromatic compound, preparation method therefor, intermediate thereof, pharmaceutical composition thereof, and use thereof
CN116196314B (en) * 2023-05-04 2023-08-15 广州市妇女儿童医疗中心 Application of RI-1 or salt thereof in preparation of medicine for preventing and treating gastrointestinal diseases
CN117603097B (en) * 2023-11-29 2024-06-18 安徽泽升科技股份有限公司 Method for rapidly preparing 2-bromo-5-nitrobenzenesulfonyl chloride

Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996031622A1 (en) 1995-04-07 1996-10-10 Oxford Gene Technology Limited Detecting dna sequence variations
WO1997010365A1 (en) 1995-09-15 1997-03-20 Affymax Technologies N.V. Expression monitoring by hybridization to high density oligonucleotide arrays
EP0834575A2 (en) 1990-12-06 1998-04-08 Affymax Technologies N.V. Methods using target nucleic acid hybridization patterns on a matrix of oligonucleotides
WO1998030883A2 (en) 1997-01-03 1998-07-16 Affymetrix, Inc. Analysis of genetic polymorphisms and gene copy number
US5837832A (en) 1993-06-25 1998-11-17 Affymetrix, Inc. Arrays of nucleic acid probes on biological chips
WO2008082856A1 (en) * 2006-12-26 2008-07-10 Pharmacyclics, Inc. Method of using histone deacetylase inhibitors and monitoring biomarkers in combination therapy
WO2014085545A1 (en) * 2012-11-30 2014-06-05 The University Of Chicago Methods and compositions involving rad51 inhibitors
WO2016094897A1 (en) 2014-12-12 2016-06-16 The Jackson Laboratory Compositions and methods relating to the treatment of cancer, autoimmune disease, and neurodegenerative disease
WO2016140971A1 (en) * 2015-03-02 2016-09-09 The Regents Of The University Of California Novel rad51 inhibitors and uses thereof
WO2016196955A1 (en) * 2015-06-04 2016-12-08 Drexel University Inhibitors of RAD52 Recombination Protein and Methods Using Same

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2782889C (en) 2009-12-17 2014-08-05 Merck Canada Inc. Aminopyrimidines as syk inhibitors
EP3652159B1 (en) 2017-07-11 2023-01-18 Cyteir Therapeutics, Inc. Substituted thiazole derivatives as rad51 inhibitors for the treatment of cancer diseases, autoimmune and neurodegenerative conditions
EP4112616A1 (en) * 2017-09-11 2023-01-04 Cyteir Therapeutics, Inc. Rad51 inhibitors
TW202104198A (en) 2019-03-12 2021-02-01 美商賽堤爾醫療公司 Rad51 inhibitors
CN114173781A (en) * 2019-03-25 2022-03-11 赛泰尔治疗公司 Combination of RAD51 and PARP inhibitors

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0834575A2 (en) 1990-12-06 1998-04-08 Affymax Technologies N.V. Methods using target nucleic acid hybridization patterns on a matrix of oligonucleotides
EP0834576A2 (en) 1990-12-06 1998-04-08 Affymax Technologies N.V. Methods using nucleic acid hybridization patterns on a matrix of oligonucleotides
US5837832A (en) 1993-06-25 1998-11-17 Affymetrix, Inc. Arrays of nucleic acid probes on biological chips
WO1996031622A1 (en) 1995-04-07 1996-10-10 Oxford Gene Technology Limited Detecting dna sequence variations
WO1997010365A1 (en) 1995-09-15 1997-03-20 Affymax Technologies N.V. Expression monitoring by hybridization to high density oligonucleotide arrays
WO1998030883A2 (en) 1997-01-03 1998-07-16 Affymetrix, Inc. Analysis of genetic polymorphisms and gene copy number
WO2008082856A1 (en) * 2006-12-26 2008-07-10 Pharmacyclics, Inc. Method of using histone deacetylase inhibitors and monitoring biomarkers in combination therapy
WO2014085545A1 (en) * 2012-11-30 2014-06-05 The University Of Chicago Methods and compositions involving rad51 inhibitors
WO2016094897A1 (en) 2014-12-12 2016-06-16 The Jackson Laboratory Compositions and methods relating to the treatment of cancer, autoimmune disease, and neurodegenerative disease
WO2016140971A1 (en) * 2015-03-02 2016-09-09 The Regents Of The University Of California Novel rad51 inhibitors and uses thereof
WO2016196955A1 (en) * 2015-06-04 2016-12-08 Drexel University Inhibitors of RAD52 Recombination Protein and Methods Using Same

Non-Patent Citations (81)

* Cited by examiner, † Cited by third party
Title
"Handbook of Pharmaceutical Excipients", 2005, PHARMACEUTICAL PRESS
"NCBI", Database accession no. 10930
"NCBI", Database accession no. 140564
"NCBI", Database accession no. 164668
"NCBI", Database accession no. 200315
"NCBI", Database accession no. 200316
"NCBI", Database accession no. 23626
"NCBI", Database accession no. 27350
"NCBI", Database accession no. 339
"NCBI", Database accession no. 403314
"NCBI", Database accession no. 5896
"NCBI", Database accession no. 5897
"NCBI", Database accession no. 60489
"NCBI", Database accession no. 7150
"NCBI", Database accession no. 7153
"NCBI", Database accession no. 7155
"Physician's Desk Reference", 2003
"Remington's Pharmaceutical Sciences", 2003
"Remington's, Pharmaceutical Sciences", MACK PUBLISHING CO.
BORCHERT ET AL., BMC CANCER, vol. 11, 2011, pages 347
CHAUDHURI ET AL., ADV IMMUNOL, vol. 94, 2007, pages 157 - 214
CHAUDHURI ET AL., NATURE, vol. 430, 2004, pages 992 - 8
CHAUDHURI; ALT, NAT REV IMMUNOL, vol. 4, 2004, pages 541 - 552
CHEN ET AL., J. BIOL. CHEM., vol. 274, 1999, pages 32931 - 32935
COLLINS ET AL., NUCLEIC ACIDS RES., vol. 29, 2001, pages 1534 - 1538
CONNELL ET AL., CANCER RES., vol. 64, 2004, pages 3002 - 3005
CROUCH ET AL., J EXP MED, vol. 204, 2007, pages 1145 - 1156
ENGELS ET AL., APPL IMMUNOHISTOCHEM MOL MORPHOL, vol. 16, 2008, pages 521 - 529
FEI HUANG ET AL: "A Small Molecule Inhibitor of Human RAD51 Potentiates Breast Cancer Cell Killing by Therapeutic Agents in Mouse Xenografts", PLOS ONE, vol. 9, no. 6, 27 June 2014 (2014-06-27), pages e100993, XP055523959, DOI: 10.1371/journal.pone.0100993 *
FELDHAHN ET AL., J EXP MED, vol. 204, 2007, pages 1157 - 1166
GODTHELP ET AL.: "Mammalian Rad51C contributes to DNA cross-link resistance, sister chromatid cohesion and genomic stability", NUCLEIC ACIDS RES., vol. 30, 2002, pages 2172 - 2182, XP002591922
GOODMAN; GILMAN: "The Pharmacological Basis of Therapeutics", PERGAMON
GREEVE ET AL., BLOOD, vol. 1010, 2003, pages 3574 - 3580
GRUBER ET AL., CANCER RES, vol. 70, 2010, pages 7411 - 7420
HANCER ET AL., LEUK LYMPHOMA, vol. 52, no. 1, January 2011 (2011-01-01), pages 79 - 84
HANSEN ET AL., INT. J. CANCER, vol. 105, 2003, pages 472 - 479
HARDIANTI ET AL., LEUKEMIA, vol. 18, 2004, pages 826 - 831
HEINTEL ET AL., LEUKEMIA, vol. 18, no. 4, April 2004 (2004-04-01), pages 756 - 62
HOCKLEY, LEUKEMIA, vol. 24, no. 5, 2010, pages 1084 - 6
HOULLEBERGHS H; GOVERDE A; LUSSEVELD J; DEKKER M; BRUNO MJ ET AL.: "Suspected Lynch syndrome associated MSH6 variants: A functional assay to determine their pathogenicity", PLOS GENETICS, vol. 13, no. 5, 2017, pages e1006765
ITO ET AL., J. GENE MED., vol. 7, no. 8, 2005, pages 1044 - 1052
KLEMM ET AL., CANCER CELL, vol. 6, 2009, pages 232 - 245
KOMORI ET AL., HEPATOLOGY, vol. 47, no. 3, 2008, pages 888 - 896
KOTANI ET AL., PNAS USA, vol. 104, 2007, pages 1616 - 1620
KOVALCHUK ET AL., J. EXP. MED., vol. 204, 2007, pages 2989 - 3001
KUMARI ET AL., EXCLI JOURNAL, vol. 7, 2009, pages 44 - 62
KUPPERS, ONCOGENE, vol. 20, 2005, pages 5580 - 5594
LEUENBERGER ET AL., MOD PATHOL, vol. 32, 2009, pages 177 - 186
LIU ET AL.: "XRCC2 and XRCC3, new human Rad51-family members, promote chromosome stability and protect against DNA cross-links and other damages", MOL. CELL, vol. 1, 1998, pages 783 - 793, XP001000712, DOI: doi:10.1016/S1097-2765(00)80078-7
LIU, M. ET AL., NATURE, vol. 451, 2008, pages 841 - 845
LIU; SCHATZ, TRENDS IMMUNOL, vol. 30, 2009, pages 173 - 181
LONGERICH ET AL., CURR OPIN IMMUNOL, vol. 18, 2006, pages 164 - 174
LONGERICH ET AL., CURR OPIN IMMUNOL, vol. 18, 2006, pages 164 - 176
MANIS ET AL., TRENDS IMMUNOL, vol. 23, 2002, pages 31 - 39
MAO ET AL., BR J DERMATOL, vol. 145, 2001, pages 117 - 122
MARUSAWA ET AL., ADV IMMUNOL, vol. 111, 2011, pages 109 - 41
MARUSAWA, INT J BIOCHEM CELL BIOL, vol. 40, 2008, pages 399 - 402
METHODS IN MOLECULAR BIOLOGY, vol. 49, 27 September 1995 (1995-09-27), pages 229 - 238
MILLS ET AL., IMMUNOL REV, vol. 194, 2003, pages 77 - 95
MOTALLEB ET AL.: "Research Journal of Applied Sciences", ENGINEERING AND TECHNOLOGY, vol. 4, 2012, pages 1888 - 1894
MURAMATSU ET AL., J BIOL CHEM, vol. 274, 1999, pages 18470 - 6
MUTO ET AL., PROC. NATL. ACAD. SCI. USA, vol. 103, 2006, pages 2752 - 2757
NAKAMURA ET AL., BR J DERMATOL, vol. 165, no. 2, 2011, pages 437 - 9
OHNISHI ET AL., BIOCHEM. BIOPHYS. RES. COMMUN., vol. 245, 1998, pages 319 - 324
OKAZAKI ET AL., J. EXP. MED., vol. 197, 2003, pages 1173 - 1181
PALACIOS ET AL., BLOOD, vol. 115, no. 22, 2010, pages 4488 - 4496
PASQUALUCCI ET AL., NAT. GENET., vol. 40, 2008, pages 108 - 112
PEREZ-DURAN ET AL., CARCINOGENESIS, vol. 28, no. 12, 2007, pages 2427 - 33
QIU ET AL., MOD PATHOL, vol. 25, no. 1, 2012, pages 36 - 45
RAMIRO ET AL., J. EXP. MED., vol. 200, 2004, pages 1103 - 1110
ROBBIANI ET AL., MOL CELL, vol. 36, no. 4, 2009, pages 631 - 41
RUSSELL ET AL., CANCER RES., vol. 63, 2003, pages 7377 - 7383
S. M. BERGE ET AL., J. PHARM. SCI., vol. 66, 1977, pages 1 - 19
SHEN ET AL., MOL. IMMUNOL., vol. 45, 2008, pages 1883 - 1892
SHIKATA ET AL., CANCER SCI, vol. 103, no. 3, 2012, pages 415 - 21
TAKATA ET AL.: "Chromosome instability and defective recombinational repair in knockout mutants of the five Rad51 paralogs", MOL. CELL BIOL., vol. 21, 2001, pages 2858 - 2866
TEBBS ET AL.: "Correction of chromosomal instability and sensitivity to diverse mutagens by a cloned cDNA of the XRCC3 DNA repair gene", PROC. NATL. ACAD. SCI. USA, vol. 92, 1995, pages 6354 - 6358
VOLPI; BRIDGER, BIOTECHNIQUES, vol. 45, no. 4, October 2008 (2008-10-01), pages 385 - 409
WHITE ET AL., AUTOIMMUNITY, vol. 44, no. 8, 2011, pages 585 - 98
XU; SCAND ET AL., J. IMMUNOL., vol. 296, 2009, pages 2033 - 6
ZHANG ET AL., HUM PATHOL, vol. 43, no. 3, 2012, pages 423 - 34

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