Specification
Field of the invention
The present invention relates to novel compounds of Formula I, their pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers including R and S isomers, polymorphs, prodrugs, metabolites, salts or solvates thereof. The invention also relates to the processes for the synthesis of novel compounds of Formula I, their pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof. The present invention also provides pharmaceutical compositions comprising compounds of Formula I and use of compounds of" Formula 1 for treating or preventing one or more conditions that may be regulated or normalized \ i;v inhibition of dipeptidyl peptidase IV (DPP-1V).
Background of the invention
Type 2 diabetes mellitus is a multifaceted and heterogeneous metabolic syndrome, which accounts for 90-95 % of all diabetes. This disorder is rapidly emerging as a global health care problem that threatens to reach pandemic levels by 2030: the number of people with diabetes worldwide is expected to rise from 171 million in 2000 to 366 million by 2030. This increase is expected to be most noticeable in developing countries, where the number of people with diabetes is expected to grow from 84 million to 228 million.
A key component of the pathophysiology of Type 2 diabetes mellitus involves an impaired pancreatic ß-cell function which eventually contributes to decreased insulin secretion in response to elevated plasma glucose. An early defect in Type 2 diabetes mellitus is insulin resistance which is a state of reduced responsiveness to circulating concentrations of insulin and is often present years before the onset of hyperglycemia and the clinical diagnosis of diabetes. The ß-cell compensates for increasing insulin resistance by increasing insulin secretion eventually resulting in reduced H-cell mass. Consequently, blood glucose levels stay at abnormally high levels, which in the long run leads to severe health problems in these patients including, obesity, hypertension and dyslipidemia. Uncontrolled hyperglycemia can further lead to complications such as nephropathy, neuropathy, retinopathy and premature atherosclerosis.
Glucose-dependent insulin secretion is mainly promoted by incretins, predominantly glucose-dependent insulinotropic peptide (G1P) and glucagon-like peptide 1 (GLP-1) (7-36). These gut peptides are released from the gastrointestinal tract in response to nutrition ingestion and promote nutrient assimilation via potentiation of glucose dependent insulin secretion, liased on its physiological profile, the actions of GLP-1 (7-36) are useful for lowering blood glucose in subjects with Type 2 diabetes mellitus and thus have strong potential as chronic therapies for diabetes. Studies in which Type 2 diabetic patients have been infused with GLP-1 have demonstrated efficacy in normalizing both fasted and post-prandial glycemia. However. GLP-1 (7-36) has been shown to have a short half-life in vivo (about 1.5 min) as it undergoes rapid amino terminal (His-Ala) degradation by dipeptidyl peptidase (DPP-IV). DPP-IV is a member of the s9b family of serine peptidases.
(Formula Removed)
A large number of DPP-IV inhibitors have been described in the art. For example. PCT publications WO-199819998, WO-2000034241. WO-2006127530, US patents US 6,110.949. US 6,011,155, US 7,169,806 and Japanese publication JP-2005139107 disclose cyanopyrrolidines as DPP-IV inhibitors. PCT publication WO-2004 101514 discloses cyanofluoropyrrolidines having DPP-IV inhibitory activity. US publications US 20006110949. US 20006107317 and PCT publication WO 199961431 disclose cyanothiazolidines as DPP-IV inhibitors. Aminopiperidine derivatives ha\e been disclosed in, for example, PCT publications Vv()-200605806 1, WO-2006039325. WO-2006058064. Others are pyrrolidine, thiazolidine, pipenuline. or pyridine derivatives (see for example WO-2006116157, WO-2005120494, WO-03084940. WO-2006062063. WO-2005042488). Still others are xanthine and purine derivatives (see tor example PCT publications WO-2004018467. WO-2004018469).
y5-amino acid based DPP-IV inhibitors have been disclosed in PCT publications, for example, WO-2004043940, WO-2005044195, WO-2006009886, WO-2006023750, WO2006039325, WO-2003004498, WO-2005116029, WO-20051 13510. WO-2006097175. WO-2005120494. WO-2005121131, WO-2005123685, WO-2005040Q95 WO-2007063928, WO-2007054577. WO-2007053819, WO-2006081151, WO-2004085378 and US patents such as US 7,259,160, US 7,101,871 and US 7,208,498.
The present invention is directed to a class ofH-amino acid based DPP-IV inhibitors using no\el heterocycles, structurally unrelated to any DPP-IV inhibitors known so far.
Although a number of DPP-IV inhibitors have been described in the art, nonetheless, a need still exists for new DPP-IV inhibitors that have better half-life, advantageous potency, stability, selectivity, less toxicity and/ or better pharmacodynamics properties. There is a need for DPP-IV inhibitors that can increase the amount of circulating GLP-I over prolonged period of time, thus leading to better control of diabetes related complications. In this regard, a novel class of DPP-IV inhibitors is provided herein.
Summary of the invention
The present invention relates to the compounds of Formula I, their pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers including R and .V isomers, polymorphs, prodrugs, metabolites, salts or solvates thereof, wherein:
(FORMULA REMOVED)
AT represents aryl which may be phenyl; which mav further be unsubstituted or may be optionally substituted at any available position by one or more substitucnts selected from but not limited to halogen, CN, hydroxyl, NH2, C>-\2 alkyl or C i-,-. alkoxv;
R1 is selected from the group consisting of but not limited to (CH2)nCONRaRb, (CH^COOR", (CH2)hNR'Rb, (CH2)nNR"CORb, (CH2)nC(-L)Ra (wherein L is O or S), (CH2)„OR" (wherein each methylene group may be substituted by one or more halogen atoms), -(CO)Ra, -(CO)NR"Rb, hydrogen, Cn2 alkyl, C2.|2 alkenyl. d ,-> alkynvl, Cn? haloalkyl, C?-l? haloalkenyl. Cri2 haloalkynyl, C3_8 cycloalkyl, heterocyclyl. aryl heteroaryl. (CH2)„-cycloalkyl, (CH2)n-heterocyelyl, (CH2)n-aryl, (CH2)n-heteroaryl, each of which may be optionally substituted at any available position by one or more substituents selected from but not limited to hydrogen, halogen. CN. Crr alkyl, C2.i2 alkenyl, C2.|2 alkynyl, C|-P alkoxv, Crp haloalkyl, C\-]? haloalkoxy. C?-]? haloalkenyl, C2-12 haloalkynyl, Cri2 alkylcarbonyl. C,-M alkoxycarbonyl, oxo. -ORa, -SRa, -N02. -NR"Rb, N(RaXCO)Rb, N(R*XCO)ORb, N(RaXCO)NRuRb. -(C'O)R'. -(CO)NRaRb, -0(CO)RJ, -0(C())NR"Rh. -COOR", Cj-g cycloalkyl, S(0)mR\ S02NRaRh; cycloalkyl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc; aryl which may be optionally substituted at any available position by one or more substitucnts independently selected from Rc or Rc; heteroaryl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc; or heterocyclyl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc';
R2 and R3 together represents a single oxygen or sulphur atom which is linked to the dia/epmc
ring by a double bond; or R1 and R2 together forms a double bond in the dia/epine ring and R3 represents the group -NRaRb; or R1 and R' together with the nitrogen atom to which R1 is attached forms a heterocyclic or heteroaryl ring which mav additionally contain from one to three heteroatoms independently selected from (). S and N; the rjng formed may optionally be substituted with one or more substituents selected from R" or R' and R2 represent hydrogen or a double bond;
R4 and R5 are independently selected from the group consisting of hydrogen, halogen, CN, Cni alkyl, C2.i2 alkenyl, C2.12 alkynyl, C|-|2 alkoxy. C'ri2 haloalkyl, Cn2 haloalkoxy. C2-i2 haloalkenyl. C2-i2 haloalkynyl, C,-,2 alkylcarbonyl, C,-12 alkoxycarbonyl, -OR", -SRa, -N02, -NRaRb, N(Ra)(CO)Rb, N(R'XCOpRb, N(R")(CO)NRaRb, -.p alkenyl, C;_p alkynyl. Cri ■ alkoxy. C1-12 alkylcarbonyl, CV12 alkoxycarbonvl. (,.s cycloalkyl, (VP haloalkyl. (VP haloalkoxy. C-p haloalkenyl, aryl, heterocyclyl, heteroaryl, (CH:)n-aryl, (CHiJn-heterocyclyl, (CH2)n-heteroaryl, (CH2)„-cycloalkyl, oxo, -CN, OR". -NO, -NR"R'", N(R"XC0)RI(I. N(R'xn)K)R10, N(R9XCO)NR9R10, -C(=L)R" (wherein I. is O or S). -(CO)NR''R'", -0(CO)R", -0(CO)NR"R'", -COOR9, -SR9, SCO^R9, S02NR9R10; SO,H. NHSO:R". P(0)RV°; or R" and Rb may be joined
together along with the nitrogen atom to which they are attached to form a heterocyclic or heteroaryl ring which may additionally contain from one to three heteroatoms independently selected from O, S and N, the ring formed may optionally be substituted with one or more substituents selected from hydrogen, halogen. C,-p alkyl. CVp alkenyl. (Vp alkynyl. C,-p haloalkyl, C2-p haloalkenyl, CVp haloalkynyl. ( :x cycloalkyl. heterocyclyl. aryl. heteroaryl. (CH2)n-cycloalkyl, (CH2)n-heterocyclyl, (CH>)„-aryl. (CH2)n-heteroaryl, Cn? alkylcarbonyl. Cri2 alkoxycarbonyl, oxo, CN. -OR9, -CF3. -OCK CH.CF,. CI\CF,, -N02, -NR9R10, N(R9XCO)R'", N(R9XCOK>R10, N(R9XCO)NR9R10. -C( l.)R" (wherein 1. is O or S), - alkoxy, or K and R1 may he joined together to torm a heterocyclic or heteroaryl ring which may contain from one to three heteroatoms independently selected from (). S and N, which may optionally be substituted with one or more substituents independently selected from Rc or Rc;
m is 1 or 2;
n is 1, 2, 3 or 4;
r is 1, 2, 3 or 4.
Another aspect of the invention provides the processes for the preparation of the novel
compounds of Formula I, their pharmaceutical^ acceptable derivatives, tautomeric forms,
stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof.
A further aspect of the present invention pro\ides pharmaceutical compositions, containing
compounds of Formula 1. their pharmaceutical^ acceptable derivati\es. tautomeric I'i'inis.
stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof in combination with one or
more pharmaceutically acceptable carriers).
Another aspect of the present invention is the use of the compounds of Formula I for the prophylaxis, amelioration and/or treatment of one or more condition(s)/disease(s)/ disorders) that may be regulated or normalized via inhibition of DPP-IV.
Yet another aspect of the invention is to provide methods of using the compounds of formula I of the present invention or compositions comprising the compounds of Formula 1 for the prophylaxis, amelioration and/or treatment of disease(s)/ disorders) mediated by FJPP-IV which comprises administering to a subject in need thereof the compounds of Formula I or compositions comprising a pharmaceutically effective amount of the compounds of Formula I.
A further aspect of the present invention is the use of a compound of formula I for the manufacture of a medicament for the prophvlaxis, amelioration and/or treatment of one or more condition(s)/disease(sy disorders) mediated bv DPP-I V in a subject in need thereof.
The present invention also encompasses prodrugs and active metabolites of the compounds of the Formula I.
Other aspects of the invention will be set forth in the description which follows, and in part will be apparent from the description, or mav be learnt bv the practice of the invention.
Detailed description of the invention
The present invention relates to the compounds of Formula I, their pharmaceutical!) acceptable derivatives, tautomeric forms, stereoisomers including R and .V isomers, polymorphs. prodrugs, metabolites, salts or solvates thereof, wherein:
(Formula Removed)
Ar represents aryl which may be phenyl: which mav further be imsubstituted or ma) be optionall) substituted at any available position by one or more substituents selected from but not limited to halogen, CN, hydroxyl, NH2. Cri2 alky! or ( rP alkow:
R' is selected from the group consisting ol but not limited to (CH2)nCONRuRb. (CH2)nCC)()Ra. (CH2)„NR'Rb, (CH2)„NR"CORb, (CH2)„C( I)R" (wherein L is O or S). (CH2)nORd (wherein each methylene group may be substituted b> one or more halogen atoms), -{CO)R\ -(CO)NRaRb, hydrogen, Cn; alkyl, C2.P. alkenyl. ('. r alkynvl. Cri? haloalkyl, C2-i? haloalkenyl. C'-.-]2 haloalkynyl, C3.g cycloalkyl, hcterocyclvl. arvl. hcteroaryl. (C'H;)„-cycloalkyl. (CHiJn-heterocvclyl. (CH2)„-aryl, (CH2)n-heteroaryl, each of which mav be optionally substituted at any available position by one or more substituents selected from but not limited to hydrogen, halogen. CN. (Vi?
alkyl, C2-12 alkenyl, C2.\2 alkynyl, Cr\?. alkoxy. CrP haloalkyl. CrP haloalkoxy. C\-P haloalkenyl, C2-12 haloalkynyl, Crn alkylcarbonyl. CrP alkoxycarbonyl, oxo, -ORa, -SRa, -N02. -NRdRb, N(R,XCO)Rb, N(RaXCO)ORb, N(Ra)(CO)NR''R'', - be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc;
R2 and R3 together represents a single oxygen or sulphur atom which is linked to the dia/epine
ring by a double bond; or R1 and R" together forms a double bond in the diazepine ring and R3 represents the group -NR"Rb; or R1 and R' together with the nitrogen atom to which R1 is attached forms a heterocyclic or heteroaryl ring which may additionally contain from one to three heteroatoms independently selected from (). S and IS: the ring formed may optionally be substituted with one or more substituents selected from K' or RL and R represent hydrogen or a double bond;
R4 and R5 are independently selected from the group consisting of hydrogen, halogen, CN, Cn - alkyl. C1.12 alkenyl, C2-12 alkynyl, CrP alkoxy. (' , haloalkyl. Crp haloalkoxy, d-P haloalkenyl. ('-,.-haloalkynyl, C1-12 alkylcarbonyl, C,-P alkoxy earbonyl. -ORJ, -SR', -N02. -NR"Rh, N(R')(( OJR1'. N(RaXCOPRb, N(RaXCO)NR'Rb, -(CO)R'. -(C'O)NR'R1'. -0(CO)R1'. -0(C())NR"Rh. -COOK1. C;x cycloalkyl, S(0)mR\ S02NRaRb ; cycloalk\l \shich ma\ be optionally substituted at an> axailable position by one or more substituents independently selected from R1^ or RL ; aryl which ma> be optionally substituted at any available position b\ one or more substituents independently selected from Rc or Rc; heteroaryl which may be optionalK substituted at any available position by one or more substituents independently selected from Rc or Rc; or heterocyclyl which may be optionally substituted at any available position by one or more substituents independently selected from R1" or
RL; R6 and R7 are independently selected from the group consisting of hydrogen, halogen. CN, CPi' alkyl, C2-12 alkenyl, C2-12 alkynyl, C|-|2 alkoxv. CrP haloalkxl. CrP haloalkoxy. C\-P lialoalken> I. C -i: haloalkynyl, C,-12 alkylcarbonyl, C™ alko\\carboml. -OR1. -SR'1, -NOp -NR'R1'. N(R'KCO)R". N(R'XCO)ORb, N(R"XCO)NR"Rb, -(CO)R'. -(('OJNR-R1', -(XCX))Rd, -0(CO)NR,,Rb. -C(K)K'. C, s cycloalkyl, S(0)„,R", S02NR"Rb : cycloalksl s^hich ma\ be optionally substituted at any axailable position by one or more substituents independently selected from R1" or Rl : aryl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc; heteroaryl which may be optionally substituted at any available position by one or more substituents independently selected from R^ or RL; or heterocyclyl which may be optionally
substituted at any available position by one or more substituents independently selected from Rl or
Rc";
R8 is independently selected from hydrogen, halogen. CN. Crp alkyl, C|-i2 haloalkyl. C|-|2 alkoxy. Cm haloalkoxy, C2-12 haloalkenyl, Cn? alkylcarbonyl. Cn? alkoxycarbonyl, -ORa, -SRa, -CF(. -OCF3, -NO2, -NRttRb, N(R"XCO)Rb, N(R°XCO)OR1', N(R"XCO)NR'Rb, -(CO)R', -(CO)NRaRb, -0(CO)R', -0(CO)NR'Rb, -COOR8, C3^ cycloalkyl, S(0)mR", S02NRaRb; cycloalkyl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc; aryl which may be optionally substituted at any available position by one or more substituents independently selected from Rl or Rc ; hetcroaryl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rl . or heterocyclyl which may be optionally substituted at any available position by one or more substituents independently selected from RL or Rl;
R* and Rb are independently selected from hydrogen. (.Vi? alkyl. C2_i?. alkenyl, CM2 alkynyl. C'rp haloalkyl, C2-12 haloalkenyl, C?-i2 haloalkynyl, (\s cycloalkyl, heterocyclyl, aryl, heteroaryl. (CH2)n-cycloalkyl, (CH2)n-heterocyclyl. (CII.->)n-aryl. (CH2)„-heteroaryl; each of which may be optionally substituted with halogen, hydroxy I. CVi.' alkyl. CM 2 alkenyl, C2-12 alkynyl, Cri2 alkoxy. Cm alkylcarbonyl, Ci-12 alkoxycarbonyl. C\.H cycloalkyl, Ci-12 haloalkyl, Cru haloalkoxy. CV12 haloalkenyl, aryl, heterocyclyl, heteroaryl. (CH2)„-aryl, (CH2)n-heterocyclyl, (CH2)„-heteroaryl, (CH2)„-cycloalkyl, oxo, -CN, -OR", -NO: -NR,;RI(I. N(R9XCO)R10, N(R9XCOX)R'0, N(R9XCO)NR9R10, -C(=L)R9 (wherein 1. is () or S), -(CO)NR9R10, -0(CO)R9, -0(CO)NR"Rlu. -COOR9, -SR9, SCO^R9, S02NR9R10; SO,H. NHSO.R9. P(0)R9RIU; or R" and Rb may be joined together along with the nitrogen atom 10 v\fiich they are attached to lbrm a heterocyclic or heteroaryl ring which may additionally contain from one to three hetcroatoms independently selected from O, S and N, the ring formed may optionally be substituted with one or more substituents selected from hydrogen, halogen, CVi.' alkyl, CM? alkenyl, CS P alkynyl. (r!: haloalkyl, C2-12 haloalkenyl, C2-i2 haloalkynyl, CiK cycloalkyl. heterocyclyl, aryl, heteroaryl, (CH2)n-cycloalkyl, (CH2)n-heterocyclyl, (CH>)„-aryl. (CH2)„-hetcroaryl, Cr\?. alkylcarbonyl. Cn.. alkoxycarbonyl, oxo, CN, -OR9, -CF3, -(X'F, CH?.CF\ CF?CF3, -NOz, -NR9R10, N(R9XCO)R10, NCR'XCOPR10, N(R9XCO)NR9R10, -C( L)R" (wherein L is O or S), -(CO)NR9R10, -0(CO)Cr C12alkyl, -0(CO)NR9R10, -COOR9, -SR9. S(()),„R9. SO,NR9R10: S03H, -NH SO?R9, -P(0)R9RHI: the ring thus formed may further be fused with 3 to 7 membered unsaturated or saturated ring, which may contain from one to three heteroatoms independently selected from O. S or N, the fused ring may optionally be substituted with one or more substituents R' or Rc;
Rc or Rc is independently selected from the group consisting of but not limited to hydrogen, halogen. Ci-12 alkyl, C2-i2 alkenyl, C2.!2 alkynyl, C|-i> haloalkyl, C2-!2 haloalkenyl, C2-|2 haloalkynyl, C ri2 alkoxy, Cn2 haloalkoxy, C3^ cycloalkyl. heterocyclyl. aryl. heteroaryl, (CH->)„-cycloalkyl. (Clli)n-heterocyclyl, (CH2)n-aryl, (CH2)n-heteroaryI, (>|, alkylcarbonyl, Cn2 alkoxycarbonyl, CN. OR9. -
OCF3, -NO2. =NOR10, -NR9R10, N(R9XCO)R10. N(R"KC0)0R10. N(R9XCO)NR9Rin, -C(L)R9 (wherein L is O or S). -(CO)NR9R'". -O(C'0)R". -(XCO)NR"R'(1. -C(X)R". -SR". S(()|M1R". S02NR9R10; SOjH, NH SO, R9, P(0)R9R".
R9 and R10 are independently selected from lndrogen. tVr alkyl. (Y|., alkcnyl, ('->_i> alkvnyl. Cm haloalkyl, C2-12 haloalkenyl, C3^ cycloalk\l. heterocyclyl. aryl, hetcroaryl, (CH:)„-cycloalk\l. (CH2)n-heterocyclyl, (CF^Xraryl, (CH2),,-licteroaryl. each ol" which may be optionally substituted with halogen, hydroxyl or C|-6 alkoxy, or Rand R " may be joined together to form a heterocyclic or heteroaryl ring which may contain from one to three heteroatoms independently selected from O, S and N, which may optionally be substituted with one or more substituents independently selected from Rc or Rc';
m is 1 or 2;
n is 1, 2, 3 or 4;
r is 1, 2, 3 or 4.
One embodiment of the present invention provides compounds of Formula la. wherein
(Formula Removed)
r, Ar, R1, R2, R3, R4, R5, R6, R7 and R8 arc as defined herein; their pharmaceutical ly acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prcxlrugs, metabolites, salts or solvates thereof.
Another embodiment of the present invention provides compounds of Formula lb. wherein
(Formula Removed)
R2 and R3 together represent a single oxygen which is linked to the diazepine ring by a double bond; r, Ar, R\ R\ R5, R6, R7 and R8 are defined herein; their pharmaceutical ly acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof. Another embodiment of the present invention provides compounds of Formula Ic, wherein
(Formula Removed)
R1 and R3 together with the nitrogen atom to which R1 is attached, form a heterocyclic or heteroarvl ring A, which is optionally substituted at an\ available position by one or more substitucnts independently selected from Re or Rc; R? either represents hydrogen or a double bond; r. Ar. R'. R\ R6, R7, R8, Rc and Rc' are as defined herein: their pharmaceutical^ acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof.
Another embodiment of the present invention provides compounds of Formula Id, wherein
(Formula Removed)
R1 and R3 together with the nitrogen atom to which R1 is attached, form a 5-mcinbered heteroc>clyl or
heteroaryl ring A wherein X, Y and Z are independently selected from the group consisting of N and
CH, die ring A is optionally substituted at any a\ailable position by one or more substituents
independently selected from R1" or R^ ; R represents a double bond; r, Ar, R1. R\ R'\ R . Rx. R' and
Rcare as defined herein; their pharmaceutically acceptable derivatives, tautomeric forms,
stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof.
Another embodiment of the present invention pro\ ides compounds of Formula Ie, wherein
(Formula Removed)
ring A is optionally substituted by Rc; R7 represents a double bond; r, Ar, R1, R\ R", R7, R8 and Rl are
defined herein; their pharmaceutical^ acceptable derivati\es. tautomeric forms, stereoisomers.
polymorphs, prodrugs, metabolites, salts or solvates thereof.
Still another embodiment of the present invention provides compounds of Formula If, wherein
(Formula Removed)
ring A is optionally substituted by one or more substituents independently selected from Rc or RL; R2 represents a double bond; r, Ar. R4, R\ R('. R\ R8. Rc and Rc' are defined herein; their pharmaceutical^ acceptable derivatives, tautomeric tonus, stereoisomers, polymorphs, prodrugs. metabolites, salts or solvates thereof.
Another embodiment of the present invention provides compounds of Formula Ig, wherein
(Formula Removed)
ring A is optionally substituted by Rc; R2 represents a double bond; r. Ar, R1, R\ R6. R', R8 and R^ are defined herein; their pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof.
Another embodiment of the present invention provides compounds of Formula Ih, wherein
(Formula Removed)
ring A is optionally substituted by one or more substituents independently selected from Rc or Rc; R2 represents a double bond; r, Ar, R4, R\ R", R . R\ R" and Rc arc defined herein: their pharmaceutically acceptable derivatives, tautomeric tonus, stereoisomers, polymorphs, prodrugs. metabolites, salts or solvates thereof.
In another embodiment of the compounds of the present invention, Ar is phenyl which is unsubstituted or substituted with 1-5 substituents independently selected from the group consisting of fluoro, bromo and C1-12 alkyl.
In a further embodiment of the compounds of the present invention, it is preferred that Ar is selected from the group consisting of 2.4.5-tnlluorophen\l. 2-fluorophenyl. 3,4-difluorophen\ I and 2,5-difluorophenyl.
In another embodiment of the compounds of the present invention, it is preferred that R* is selected from the group consisting of hydrogen, halogen. CYr alkyl. (Yi: alkoxy and Cri.' alkoxycarbonyl.
In a further embodiment of the compounds of the present invention, it is more preferred that R is selected from the group consisting of hydrogen, fluoro, chloro and methoxy.
In a further embodiment of the compounds of the present invention, it is still more preferred that R is independently selected from hydrogen and fluoro.
In another embodiment of the compounds of the present invention, R1 is selected from the group consisting of hydrogen, (CH2)„CONRuR'. -(C'OJNR-'R1'. -(CO)Ru, (CI h)nCOOR", (CH1)„NR"Rb, (CHzyMCOR", (CHACC-LJR1' (wherein 1. is () or S). C ,-,. alk\l. C..,-, alkenyl. C,-,. haloalkx I. (',- haloalkoxy, C2-12 haloalkenyl, C2.\? alkenyl. (Y c\cloalk\l. heteroeyclyl. aryl. hcteroarxl. (C'll.),,-cycloalkyl, (CH2)n-heterocyclyl, (CH2)„-ar>l. alkylcarbonyl, CYi' alkoxycarbonyl. oxo, -OR", -SR\ -N02,-NR"Rb, -N(R'XCO)Rb, -N(RJKCOK)R1'. -N(Ra)(CO)NRilRb, -(CO)Ra. -(CO)NR'Rh. -CKCCOR*, -0(CO)NR,Rb, -COORa, C-,_s cycloalkyl. heteroaryl. heteroeyclyl.
In a further embodiment of the compounds of the present invention, it is preferred that R1 is selected from the group consisting of: hydrogen. (Yr alkyl. CY12 alkenyl. CM: alkynyl, Cru
haloalkyl, C,-12 haloalkoxy, C2-12 haloalkenyl.
(Formula Removed)
Another embodiment of the present invention provides compounds of the Formula k\ Formula Id, Formula le. Formula If, Formula Ig and lormula Ih wherein the ring A is substituted with one or more Rc and Rc independently selected from the group consisting of halogen. alkenyl, C2.12 alkynyl, Cri2 haloalkyl, C>|. haloalkenyl. C'.-i.- haloalkynyl, Cr|. alko.w. C i-P haloalkoxy, C3.8 cycloalkyl, heterocyciyi. an. I. hcteroar\l. (C'll?)M-eycloalkyl, (CH:)M-heterocveKI, (CH2)„-aryl, (CHzXrheteroaryl, C1-12 alk>lcarbon\l. C rr alkoxvcarbonyl, -CN. OR". -OCF,. -NO, =NOR10, -NR9R10, -N(R9XCO)R10, -NCRV'OJOR'". -N(R9XCO)NR9R10, -C( L)R'' (wherein L is O or S), -(CO)NR9R10, -0(CO)R9, -0(CO)NRV", -C(X)R9, -SR9, -S(0)mR9, -S02NR9R10; -SO,H, -NHSO2R9 and P(0)R9R10 , where R9 and R1" are as defined herein;
In a further embodiment of the compounds of the Formula Ic, Formula Id. Formula le. Formula If, Formula Ig and Formula Ih, it is preferred that ring A is substituted with one or more R' and Rc independently selected from the group consisting of hydrogen, (V12 alkyl. alkeml. alkynyl, C3^cycloalkyl. phenyl, -CH2F, -CHI -. -CI .. COOII.-CONIU -CI I -OCI I,. C'OOt |(,alk> I.
(Formula Removed)
In an embodiment of the compounds of the present invention, R and R are independent^ selected from the group consisting of hydrogen, halogen. (|-p alkyl. CS.p alkcnyl. C\.p alkynvl. Cpp alkoxy, Cri2 haloalkyl, Cpii haloalkoxy, C-| • haloalkenvl. C-i > haloalkynyl. -NO- cycloalkv I . aryl , heteroaryl or heterocyclyl, each of which ma\ be unsubstitutcd or substituted, at any available position, with one or more substituents independently selected from R1" or Rc.
In a further embodiment of the compounds of the present invention, it is preferred that R4 and R5 are both hydrogen.
In an embodiment of the compounds of the present invention, R6 and R are independently selected from the group consisting of hydrogen. C'ri? alkyl and C'rr alkoxy, which are unsubstituted or substituted, at any available position, with one or more substituents selected from the group consisting of halogen, hydroxyl, Crp alko\_\ and ar> I
In a further embodiment of the compounds of the present invention, it is preferred that R' and R7 are both hydrogen.
Relative to the above description of the compounds of the present invention, the following definitions apply.
Unless specified otherwise, the terms "alk\l". 'alkenyf, and "alkynyl" may be straight or branched with 1 to 12 carbon atoms. These groups ma\ further be substituted with one or more substituents selected from but not limited to. for example, halogen, hydroxyl, oxo, carboxyl. carboxyalkyl, azido, alkenyl, alkynyl, alkow. c\cloalkyl. eycloalkynyl, acyl acyloxy, aryl, heterocyclyl and heteroaryl.
The term "cycloalkyl" refers to evclic alkyl groups constituting of 3 to 20 carbon atoms having a single cyclic ring or multiple condensed rings, for example, fused or spiro systems which may optionally contain one or more olefinic bonds, unless otherwise constrained by the definition. Such cycloalkyl groups include, by way of example, single ring structures, for example, cyclopropyl. cyclobutyl, cyclopentenyl, cyclohexyl, cycloochl. and the like, or multiple ring structures, for example, adamantyl. and bicyclo[2.2.1J heptane, or c\clic alk\l groups to which is fused an ar\l group, for example, indane and the like. C\cloalk>l groups may further be substituted with one or more substituents selected from but not limited to. for example, halogen, hydroxyl. oxo, carboxv.
carboxyalkyl, azido, alkenyl. alkynyl. alkoxy. cycioalkyl. cycloalkyny I. acyl acyloxy. ar>l. heterocyclyl, heteroaryl.
The term "alkoxy" denotes the group O-alkyl wherein alkyl is the same as defined above.
The term "aralkyl" refers to alkyl-aryl linked through alkyl (wherein alkyl is the same as defined above) portion and the said alkyl portion contains carbon atoms from 1-6 and the ar> 1 is as defined herein, after. The examples of aralkyl groups include benzyl and the like.
The term "aryl" herein refers to a carbocyclic aromatic group, for example phenyl or naphthyl ring and the like optionally substituted with one or more substituents selected from but not limited to, for example, halogen, hydroxyl, alkyl, alkenyl. alkynyl, cycioalkyl, alkoxy, acyl, arylox>, CFi. COORd (wherein Rd can be hydrogen. alkyl. alkenyl. cycioalkyl, aralkyl, heterocyclylalkyl or heteroarylalkyl), cyano, nitro, carboxy, heterocyclyl, lieieroaryl. heterocyclylalkyl or hcteroary lalkyl. « The aryl group may optionally be fused with cycloalky I group, wherein the said cycioalkyl group may optionally contain heteroatoms selected from (>. N and S.
The term "aryloxy" denotes the group O- aryl wherein aryl is as defined above.
The term "heteroaryl" unless and otherwise specified refers to an aromatic ring structure or a bicyclic aromatic group with one or more heteroatom(s) independently selected from N. () and S and optionally substituted at any available position by substituent(s) selected from but not limited to halogen, hydroxyl, alkyl, alkenyl, alkynyl, cycioalkyl. acyl. carboxy, aryl. alkoxy, aralkyl, cyano. nitro, heterocyclyl, or heteroaryl. Examples of heteroaryl groups include oxazolyl, imidazolyl, pyrrolyl. 1.2.3.-triazolyl, 1,2,4-triazolyl, tetrazolyl, thia/olyl. oxadia/olyl. benzoimidazolyl, thiadiazolyl. pyridinyl, pyridazinyl, pyrimidinyl, thienyl, isoxazolyl, triazinyl. furanyl, ben/ofuranyl, indolyl, benzothiazolyl, benzoxazolyl, and the like.
The term "heterocyclyl" unless and otherwise specified refers to a cyclic, bicyclic or tricyclic cycioalkyl group, fully or partially unsaturated having 5 to 10 carbon atoms; with one oi more heteroatom(s) independently selected from N. (J and S. and are optionally ben/of used or fused with heteroaryl of 5-6 ring members; the rings may be optionally substituted wherein the substituents are selected from but not limited to halogen, hydroxyl. alkyl, alkenyl. alkynyl, cycioalkyl, acyl. carboxy. aryl, alkoxy, aralkyl, cyano, nitro, heterocyclyl, or heteroaryl. l-.xamplcs of heterocyclyl groups include but are not limited to oxazolidiny I. tetrahydrofuranyl, dihydrofuranyl. dihydropyridinyl, dihydroisooxazolyl, dihydrobenzofuryl, azabicyclohexyl, dihydroindonyl, piperidinyl or pipera/inyl.
"Heteroarylalkyr refers to alkyl-heteroaryl group linked through alkyl portion, wherein the alkyl and heteroalkyl are the same as defined previously.
"Heterocyclylalkyl" refers to alkyl-hetcrocyclyI group linked through alkyl portion, wherein the alkyl and heterocyclyl are the same as defined pre\ iously.
Halogen refers to fluoro, chloro. bromo or iodn
The term "Protecting Group" or "P( i" refers to a group which is in a modified form to preclude undesired side reactions at the protected site. The term protecting group, unless otherwise
specified, may be used with groups, for example, hydroxyl. amino, carboxyl and examples of such groups are found in T.W. Greene, et al. "Protecting iiroups in Organic Synthesis." 3rd IZd. Wiley. New York, which is incorporated herein by reference. The species of the carboxylic protecting groups, amino protecting groups or hydroxxl protecting groups employed are not critical, as long as the derivatised moieties/moiety is/arc stable to conditions of subsequent reactions and can be removed without disrupting the remainder of the molecule, l-.xamples of suitable hydroxyl and amino protecting groups include but are not limited to trimethylsilyl. triethylsilyl, u-nitrobcnzyloxycarbonyl, /Miitrobenzyloxycarbonyl, /-butyldiphenylsiKl. /-butvldimethylsilyl. acetyl. trifluoroacetyl. benzyloxycarbonyl (CBz), f-butoxycarbonyl (Boc). ^-fluorenylnethylcnoxycarbonyl (l-'moe). 2.2.2-trichloroethyloxycarbonyl, allyloxycarbonyl and the like, l-.xamples of suitable carboxyl protecting groups are benzhydryl, o-nitrobenzyl, />-nitrobenzyl. 2-naphthylmethyl, allyl, 2-chloroallyl. benzyl, 2,2,2- trichloroethyl, trimethylsilyl, /-butyldimethylsilyl, /-butyldiphenylsilyl, 2-(trimethylsilyl)ethyl, phenacyl./Mnethoxybenzyl, acetonyl, p-methoxyphenyl. 4-pyridylmethyl, f-butyl and the like.
The term "therapeutically effective amount'" means the amount of a compound that, when administered to a subject for treating a disease, is sufficient to effect such treatment for the disease. The "therapeutically effective amount" will \ar\ depending on the compound, the disease and its severity, weight, physical condition and responsiveness of the subject to be treated, among other factors.
A "pharmaceutically acceptable salt" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic or organic bases and inorganic or organic-acids.
Asymmetric centres may exist in the compounds of the present invention. The compounds ol Formula I may have one or more stereogenic centres and so can exhibit optical isomerism. All such isomers including enantiomers, diastereomers. and epimers are included within the scope of this invention. Furthermore, the invention includes such compounds as single isomers (R and /or S) and as mixtures, including racemates. If desired, racemic mixtures of the compounds may be separated so that the individual enantiomers are isolated I he separation ma\ be carried out b\ methods well known in the art, such as the coupling of a racemic mixture of compounds to an cnantiomcricall\ pure compound to form a diastereomeric mixture, followed by separation of the individual diastereomers by standard methods, such as fractional crystallization or chromatography. Starting materials of particular stereochemistry may either be commercially available or may be made by the methods described herein and resolved by techniques well known in the art.
The Formula I shows the structure of compounds without any preferred stereochemistry. formula la shows me preferred stereochemistry at the carbon atom to which is attached the amino group of the P amino acid, from which these compounds are prepared.
The independent syntheses of these diastereomers or their chromatographic separations may be achieved as known in the art by appropriate modifications.
Certain compounds according to Formula I. can also exist as tautomers. which have different points of attachment of hydrogen accompanied by one or more double bond shifts. These tautomers, either separately or as mixtures, are also considered to be within the scope of the invention.
The present invention also encompasses geometrical isomers of compounds of Formula 1 and the mixtures thereof.
Particularly useful examples of the present invention include but are not limited to the compounds selected from Tables 1 to 5:
One embodiment of the present invention pro\ ides compounds of Formula lb. wherein the compounds are selected from Table 1:
(Table Removed)
One embodiment of the present invention provides compounds of Formula If. wherein the compounds are selected from Table 3:
(Table Removed)
One embodiment of the present invention pro\ ides compound of Formula Ig wherein the compounds are selected from Table 4:
Table 4
(Table Removed)
One embodiment of the present invention pro\ ides compound of Formula Ih. wherein the compound is in Table 5
Table 5
(Table Removed)
The compounds of the present invention can be prepared from /i-amino acid intermediates such as those of Formula II, wherein Ar is as defined herein and heterocyclic intermediates such as those of Formula III, wherein r. R1. R2, R\ R'. R . R". R and RK are as defined herein, using standard coupling conditions followed by deprotection of the amine protecting functionality, l-.xamplcs of standard coupling conditions include EDC | l-ethyl-3-( Vdimethylaminoprop\l)earbodiimide|/H()B 1
(1-hydroxybenzotriazole); DCC (dicyclohexyl carbodiimide). DMAP (4-dimethylaminopyridine); HATU [0-(7-azabenzotriazole-yl)-Af,A(f,A''. \ -tetramcth\luronium hexafluorophosphate: HOA I (1-hydroxy-7-azabenzotriazole); BOP |(ben/oiria/ok l-l-\ loxv )-H-/.v(dimcthvlaminc) phosphomum hexafluorophosphate]; mixed anhydride method using ethvl chloroibrmate or mcthvl chloroformute in a suitable solvent such as DMF. DCM, acetOnitrile. toluene. IMF and the like or mixtures thereof and in the presence of a suitable base such as NMM (A'-methylmorpholine). DIPFA (Y.Y-diisopropylethylamine), triethylamine and the like, hxamples of reagents used for deprotecting the amine protecting moiety will depend upon the nature of protecting group used. Hxamples of suitable amino protecting groups include but are not limited to acetyl, trifluoroacetyl, benzyloxycarbonyl (CBz), /-butoxycarbonyl (Boc), 9-fluorenvlnethylenoxycarbonyl (Fmoc), 2,2,2-trichloroediyloxycarbonyl, allyloxycarbonyl and the like. The appropriate conditions for the removal of the amine protecting groups can be readilv selected by those having well known skill in the art. Examples of reagents used for deprotecting the amine protecting moiety include but are not limited to use of acidic conditions (trifluoroacctic acid, hydrochloric acid, phosphoric acid, p-toluencsulphonic acid and the like), basic conditions (pipcridine and the like) or hydrogenation conditions (palladium on charcoal or platinum and the like). The resulting compounds may be in the form of free amine or salt depending upon the nature of the protecting group and the corresponding deprotecting reagent used. In case the deprotection results in the formation of salt, the corresponding amine can easilv be obtained by treating the salt with an appropriate base such as triethylamine. diethylisopropylamine. sodium bicarbonate, sodium hydroxide or the like.
(Table Removed)
Enantiomerically pure/? -amino acids having the Formula II may be conveniently synthesized using methods described in Tetrahedron. 1994. 32. 9517.: Eiumtio.se/ective Synthesis of fi-Amino Acids, Ed., Wiley-VCH, New York: 1997: Aldrichiniica Acta. 1994, 27:3 and Angew Chein Ini Ed Engl. 1981, 20, 798.
In particular, 3-amino-4-(2,4,5-trifluoro-phenyl)-butyric acid may be synthesized as reported in the patent application WO 2004069162 and in./. Med. Chem., 2005, 48, 141.
Compounds of Formula III can easily be prepared from compounds of Formula IV, wherein r,
1 "J 1 A i f\ 7 a
R , R~, R , R , R , R , R and R are as defined herein, by deprotecting the amine protecting group, using standard deprotecting reagents. The resulting compounds mav be in the form of free amine or salt depending upon the nature of the protecting group and the corresponding deprotecting reagent
used. Examples of reagents used for deprotecting the amine protecting moiety include but are not limited to use of acidic conditions (trifluoroacetic acid, hydrochloric acid, phosphoric acid, p-toluenesulphonic acid and the like), basic conditions (piperidine and the like) or hydrogenation conditions (palladium on charcoal or platinum and the like). The resulting compounds may be in the form of free amine or salt depending upon the nature of the protecting group and the corresponding deprotecting reagent used. In case the deprotection rcsultb in the formation of salt, the corresponding amine can easily be obtained by treating the salt with an appropriate base such as triethylamine. diethylisopropylamine, sodium bicarbonate, sodium hydroxide or the like.
(Table Removed)
Compounds of Formula IV can be easily prepared by a variety of methods familiar to those skilled in the art. Some common routes for the preparation of such compounds arc illustrated in Schemes 1 to 11.
Compounds of Formula IV. can gencralk bo svnthesisecl from compounds of Formula V. wherein r, R4, R5, R6, R7 and R8 are as delined herein [Ref: Synthesis, 2005, 18811. One convenient route for the synthesis of compounds of Formula V is outlined in Scheme 1.
(Scheme Removed)
The compounds of Formula V can be prepared starting from the appropriate 2-nitrobenzyl
bromide which is coupled with the required ester of an amino acid in the presence of a base such as DIPEA, NMM, potassium carbonate and the like and a solvent such as DMF. I 111-, ucctonitrilc and the like, resulting in the formation of compounds of formula VI, wherein r. R1. R^ and Rs arc as defined herein. The compounds of Formula VI are then protected at the amine functionality using the appropriate protecting group resulting in compounds of Formula VII. Compounds of Formula VII, wherein r, R4, R5 and R8 are as defined herein, are then converted to compounds of Formula VIII, wherein r, R4, R5 and R8 are as defined herein. b> hydrolyzing the ester moiety under basic conditions. Examples of basic hydrolyzing reagents include sodium hydroxide, lithium hydroxide, potassium hydroxide and the like, which can be used in appropriate solvents like THF/water, dioxane/water or
THF/MeOH/water. The nhro group of Formula VIII can then be reduced to the corresponding amino
compounds of Formula IX, wherein r, R4, R' and R8 arc as defined herein, by a variety of reducing
agents such as hydrogenation over an appropriate catakst such as palladium, platinum, or ruthenium
on activated charcoal or chemical methods such as reaction with lcC'li or SnCU/lK'l or Fe/NlljC "I or
NiCl2/NaBH4 or Fe/HCI familiar to those skilled in the art. Compounds of Formula IX containing an
amine and carboxylic acid functionality are then cyclized under standard coupling conditions, for
example, using EDC [l-ethyl-3-(3-dimethylaminopropyl)carbodiimideJ/HOBT (1-
hydroxybenzotriazole) or DCC (dicyclohex> I carbodiimide), DMAP (4-dimethylaminopyridine) or HATU [O^T-azabenzotTiazole-ylVMA^A^NMetramethyluronium hexafluorophosphatc or HOAT (1-hydroxy-7-azabenzotriazole ) or BOP [(ben/otria/.ol\l-l-\lo\y)-H7\(dimeth\lamine) phospliomuin hexafluorophosphatej or by mixed anludndc method using ethyl chloroformate or mellnl chloroformate in a suitable solvent such as DM1 . IX M. acetonitrile, toluene or Till- and the like or mixtures thereof and in the presence of a suitable base such as NMM (/V-methylmorpholine). DIPF.A (A'.A'-diisopropylethylamine) or triethylamine. to afford compounds of Formula V.
(Formula Removed)
A convenient route to prepare compounds of Formula XVII, wherein R4 and R5 are as defined herein, is as described in Scheme 2.
The compounds of Formula XV11 can be prepared starting from 2-amino- l.^-dilliioroben/oie acid, which is reduced using appropriate reducing agents such as lithium aluminium hydride or boiane dimethylsulphide or mixed anhydride/sodium borohydridc in a suitable solvent such as diethyl ether or THF or DCM or mixtures thereof, to obtain compounds of Formula X. The compounds of Formula X are then protected using amine protecting reagents such as bcn/ylchlorofonnate or di-/tv7-but\l dicarbonate or Fluorenylmethyloxycarbonyl chloride (Fmoc-CI) in a suitable solvent such as DCM or dioxane or THF, resulting in compounds of Formula XI. The compounds of Formula XI are then chlorinated under standard chlorinating conditions using thionyl chloride or POCI3 or PCI;, resulting in the compounds of Formula XII, which are further coupled with the suitably protected amino acid using an appropriate base such as DIPKA or tricthylamine or potassium carbonate or cesium carbonate or sodium carbonate and the like in an appropriate solvent to afford compounds of Formula XIII, wherein R4 and R5 arc as defined herein I he compounds o\' Formula XIII are then again protected at the amine moiety using a different protecting group than the earlier used to protect compounds of Formula X, to form compounds of Formula XIV, wherein R4 and R5 are as defined herein, which are then hydrolysed under basic conditions resulting in compounds of Formula XV. wherein R4 and R3 are as defined herein. F.xamples of basic hydrolyzing reagents include sodium hydroxide, lithium hydroxide, potassium hydroxide and the like, which can be used in appropriate solvents like THF/water, dioxane/water or I HF/MeOI l/water. Compounds of Formula XV are selectively deprotected at one of the amine moieties to afford compounds of Formula XVI, wherein R4 and R5 are as defined herein, which are then subjected to internal cyclization to form compounds of Formula XVII. The appropriate conditions for the selective removal of one of the amine protecting groups can be readily selected by those having well known skill in the art. Examples of standard cyclization conditions include EDC f l-eth\l-.V(3-ilimcth\laminopropyl)carbodiimidc|/IIOB I (1-hydroxybenzotriazole); DCC (dicyclohe\\ I carbodnmidc). DMAP ( I-1 -yloxy)-/r/'.s(dimethylamine) phosphonium hexafluorophosphate]; mixed anhydride method using eth> I chloroformate or meth>l chloroformate in a suitable solvent such as DMF, DCM, acetonitrile. toluene, FHF and the like or mixtures thereof and in the presence of a suitable base such as NMM (N-methylmorpholine). DIPHA (Y./V-diisopropylethylamine), triethylamine and the like.
(Scheme Removed)
The compounds of the Formula XX. wherein r. R1. R\ Rs and Rc arc as defined herein, can be prepared from compounds of Formula V, as described in Scheme 3. The compounds of Formula V are converted to compounds of Formula XVIII. wherein r. R1. R1 and R are as defined herein. b\ reacting with Lawesson's reagent or phosphorus pcntasulphidc in the presence of a suitable solvent such as toluene, benzene, xylene.dioxane and the like or mixtures thereof under suitable heating conditions [Ref: J. Org. Chem., 1964, 29. 2311. I he compounds of Formula XV111 are then reacted with compounds of Formula XIX, wherein R' is as defined herein, under heating conditions in an
appropriate solvent such as benzene or toluene or DMSO or mixtures thereol" to give compounds of Formula XX [Ref: reported in patent application no. WO 96209411.
(Scheme Removed)
The compounds of Formula XIX are either commercially available or can be prepared by
those skilled in the art. One convenient route lor the preparation of such compounds is described in
Scheme 4. Starting with the commercialK available /c/v-butyl carbamate and reacting it with the
appropriately substituted acid or acid chloride, under standard coupling conditions, compounds of
Formula XXI, wherein Rc is as defined herein, can be prepared, which can then be deprotected under
acidic conditions such as using trifluoroaeeiie acid or by passing hydrochloride gas or />■
toluenesulphonic acid and the like, resulting in compounds of formula XIX or their respecti\e salts.
Examples of standard coupling conditions include HDC | l-eth\ !-.?-(.>-
dimethylaminopropyl)carbodiimide]/HOB1 (1 -hydroxyben/otria/ole); IX'C (dicyelohexyl carbodiimide), DMAP (4-dimethylaminop>ndinc): HAIU |0-(7-azaben/otria/ole-yl)-A'.A.A .A"-tetramethyluronium hexafluorophosphate: HOAI (l-h>droxy-7-azabenzotria/.ole); BOP [(benzotriazolyl-l-yloxy)-fris(dimetJiylamine) phosphonium hexafluorophosphate]; mixed anhydride method using ethyl chloroformate or methyl chlorofomiate in a suitable solvent such as DMF, DCM, acetonitrile, toluene, THF and the like or mixtures thereof and in the presence of a suitable base such as NMM (N-methylmorpholine), DIPEA (A',.V-diisopropylethylamine), triethylamine and the like.
(Formula Removed)
The compounds of Formula XXIX. wherein r. R" and R8 are as defined herein, can be prepared following the procedure described in Scheme 5. starling from the appropriately substituted 2-nitro benzyl bromide, which can be con\ertcd to the corresponding a/ides using sodium a/ide in a suitable solvent like DMF or THF or toluene under heating conditions to form compounds of Formula XXII, wherein r and R are as defined herein, which are then reduced under neutral reducing conditions using triphenyl phosphine in IIII/water, resulting in the formation of corresponding amines of Formula XXIII, wherein r and Rs are as defined herein. The amine functionalitv of compounds of Formula XXIII was protected with a suitable protecting group such as benzylchloroformate or di-ter/-butyl dicarbonate or Fluorenylmethyloxycarbonyl chloride (Fmoc-CI) in a suitable solvent such as DCM or dioxane or 11 IF. to afford compounds of Formula XXIV. wherein r and R8 are as defined herein. The nitro group of compounds of Formula XXIV can then be reduced to the corresponding amino groups of Formula XXV. wherein rand R8 arc as defined herein, by a variety of reducing agents such as hydrogenalion over an appropriate catalyst such as palladium. platinum, or ruthenium on activated charcoal or chemical methods such as reaction with I cC I or SnCl2/HCl or Fe/NF^Cl or NiCl2/NaBH4 familiar to those skilled in the art. The amino compounds of Formula XXV are further subjected to dia/.oti/ation reaction using sodium nitrite in the presence of acetic acid/water, followed by reaction with sodium a/ide. to afford compounds of Formula XXVI. wherein rand R8 are as defined herein. I'he compounds of formula XXVI are ihen reacted with propargyl bromides of the formula XXVII. wherein R' is as defined herein, resulting in the formation of compounds of Formula XXVIII, wherein r. R8and RL are as defined herein, which upon heating in a suitable solvent like benzene, toluene, xylene and the like or mixtures thereof, resulted in the formation of compounds of Formula XXIX |Rcf: Org.l.ett., 2008, 10, 1617|. The substituted propargyl bromides of the formula XXVII, which are not commercially available can be prepared by those skilled in the art by following procedures well documented in literature. Kxamples of such procedures include but are not limited to usnm Sonogitshira reaction of proparg\ I alcohol with the appropriate halides, followed by conversion ol alcohol to bromide |Ref: Tel. Leu. 1975, 50. 446"?../. Org. Chem., 1993, 55, 4716; WO 95/244001.
The compounds of Formula XXXI. wherein r. R1. R" and Rx are as defined herein, can be prepared following the procedure described in Scheme 6. starting from compounds of Formula XVIII which upon reaction with 2-aminoethanol result in the formation of compounds of Formula XXX. wherein r, R4, R5 and R8 are as defined herein. The compounds of Formula XXX are then subjected to Swern Oxidation followed by internal cyclization to give the compounds of Formula XXXI, wherein r, R4, R'and R8 are as defined herein. [Ref: WO 96/20941. WO 96/23790, F.P1 183243].
(Scheme Removed)
The compounds of Formula XXXIII, wherein r. R1. R5 and R8 are as defined herein, can be prepared by following the procedure as described in Scheme 7. The compounds of Formula V (where R6 , R7 = H), are reacted with diethylchlorophosphate using various bases such as sodium hydride. Lithiumdiisopropylamide, DBU, potassium carbonate and potassium /e/7-butoxide. in a suitable solvent like THF or DMF, to afford compounds o\' Formula XXXII. wherein r. R1. R' and R* au- as defined herein. The compounds of Formula XXXII are then reacted with ethylisocvanoacetatc using potassium ter/-butoxide as a base, using THF as a solvent, to give the compounds of Formula XXXIII [Ref: J. Med Chem., 2008, 57, 4370 and references mentioned therein].
The compounds of Formula XXXVI, wherein r. Ar. R1. R\ R8. R" and Rh arc as defined herein, can be prepared by following the procedure as described in Scheme 8. The compounds of formula XXXIII, wherein r, R4, R5 and R8 are as defined herein, were deprotected at the amine functionality and then coupled with compounds of Formula II, under standard coupling conditions, resulting in compounds of Formula XXXIV, wherein r. Ar. R1. Rs and R8are as defined herein. Examples of standard coupling conditions include EDC | l-ethyl-3-(3-dimethylaminopropyl)carbodiimideJ/HOBI (1-hydroxybenzotriazole); DCC (dicyclohexyl carbodiimidc). DMAP (4-dimethylaminopyridine); HATU [0-(7-azabenzotria/ole-yl)-.V^\',;V .A -tetrametln luronium hexafluorophosphate: IIOAI (1-hydroxy-7-azabenzotriazole); BOP |(ben/.olriay.ol> 1-1 -yloxy)-/r/.v(dimethylamine) phosphoimini hexafluorophosphate]; mixed anhydride method using eth\l chioroformate or methyl chlorofomiatc in a suitable solvent such as DMF, DCM, acetonitrile, toluene, THF and the like or mixtures thereof and in the presence of a suitable base such as NMM (A'-methylmorpholine). DIPHA (A.A-diisopropylethylamine), triethylamine and the like. 1 he compounds of Formula XXXIV were then
hydrolysed under basic conditions using appropriate base to gi\c the compounds of lormula WW. wherein r, Ar, R4, R5 and R8are as defined herein, which were further coupled with appropriate amines under standard coupling conditions, resulting in the formation of compounds of Formula XXXVI. Examples of basic hydrolyzing reagents include sodium hydroxide, lithium hydroxide, sodium methoxide and the like, which can be used in appropriate solvents like TUF/water, dioxane/water or THF/MeOH/water.
(Scheme Removed)
The compounds of Formula XXXVII. wherein r. R1, R*. R\ R6. R7 and R8 arc as defined herein, can be prepared from the compounds of lormula V b% following the procedure as described in Scheme 9. The compounds of Formula V were N-alk\lated at the amide position using appropriate halide in the presence of suitable bases like sodium hydride, potassium carbonate or cesium carbonate, in a suitable polar solvent such as DMF. THF and the like or the mixtures |Ref: US 20060148790 and J. Med Chem, 2007, 5(K 55b \\.
(Scheme Removed)
The compounds of Formula XXXIX. wherein r. R1. R\ R". R . R8. R" and R1' are as defined herein, can be prepared by following the procedure as described in Scheme 10. I he compounds of Formula XXXVII, wherein R1 is CH2COOi;t and r. R1. R\ Rr>, R' and R8 are as defined herein, can be
hydrolysed under basic conditions to afford the compounds of Formula XXXVIII, wherein r, R\ R5. R6, R7 and R8 are as defined herein, which are further coupled to various amines under standard coupling conditions to provide the compounds oi" Formula XXXIX. Examples of basic hydroly/.ing reagents include sodium hydroxide, lithium h\dro\ide. sodium methoxide and the like, which can be used in appropriate solvents like THF/water. dioxane/water or IHF/MeOH/water. 1-Aamples of standard coupling conditions include FIX | l-clhyl-3-(3-dimcth\laminopropyl)carhodiimidc|'ll()BI (1-hydroxybenzotriazole); DCC (dicyclohex\l carbodiimide). DMAP (4-dimethylaminopyridine): HATU [O^T-azabenzotriazole-yO-MMA^'./V-tetraniethyluronium hexafluorophosphate: MOAT (I-hydroxy-7-azabenzotriazole); BOP |(bcn/otriazol\ I-1 -yloxy )-/ra(dimethylaminc) phosphonium hexafluorophosphate]; mixed anhydride method using eth\l chloroformate or methyl chloroformate in a suitable solvent such as DMF, DCM, acetonitrile. toluene, THF and the like or mixtures thereof and in die presence of a suitable base such as NMM OY-methylmorpholine). DIPEA (VA-diisopropylethylamine), triethylamine and the like.
(Scheme Removed)
As shown in Scheme 11, the compounds of formula XXXIII can alternatively be coin cited to compounds of Formula XXXX, wherein r. R1, R\ and R's arc as defined herein, via basic hydrolysis followed by coupling with the appropriate amine resulting in the formation of compounds of Formula XXXXI, wherein r, R4, R5, R8, R* and R" arc as defined herein. Examples of basic hydroKzing reagents include sodium hydroxide, lithium hydroxide, sodium methoxide and the like, which can be used in appropriate solvents like THF/water. dioxane/water or TI IF/MeOH/water. Examples of standard coupling conditions include HDC | l-cthyl-3-(3-dimethylaminopropyl)carbodiimide|/lK)BT (1-hydroxybenzotriazole); OCC (dicyclohex\ I carbodiimide). DMAP (4-dimethylaminopyridine);
HATU [0-{7-azabenzotriazole-yl)-A^V,Vv"'.Ar'-tetramethyluronium hexafluorophosphate; HOAI (1-hydroxy-7-azabenzotriazole); BOP [(ben/otriazol\l-l-\loxy)-/ra(dimethylamine) phosphonium hexafluorophosphate]; mixed anhydride method using ethyl chlorotormate or methyl chloroforniate in a suitable solvent such as DMF. DCM, acetonitrile. toluene. THF and the like or mixtures thereof and in the presence of a suitable base such as NMM (A-methylmorpholinc). DIPF,A (A'.A-diisopropylethylamine), triethylamine and the like.
(Formula Removed)
Some representative examples of the novel heterocyclic intermediates synthesized using the procedures described in Schemes 1-11, include but are not limited to:
(Scheme Removed)
wnerein r is i.^oort
wherein R8 is selected from a group consisting of H, F, CI and OCH3
wherein R1 is selected from a group consisting of -CH3,
(Formula Removed)
where the moiety is selected from the group consisting of
(Formula Removed)
It is understood that, as used herein, references to the compounds of structural Formula I arc meant to also include the pharmaceutical!} acceptable salts, and also salts that are not pharmaceutical ly acceptable when they arc used as precursors to the free compounds or their pharmaceutical ly acceptable salts or in other synthetic manipulations. The compounds of the present invention may be administered in the form of a pharmaceutical^ acceptable salt. The term "pharmaceutically acceptable salt" refers to salts prepared from pharmaceutical ly acceptable nontoxic bases or acids including inorganic or organic bases -dial inorganic or organic at ids. I'lie sails ma\ be prepared during the final isolation and purification of the compounds or separately b\ making basic or acidic addition salts. Representative salts ot basic compounds of the present invention can be prepared by reacting free base form of the compound with a suitable acid, including, but not limited to acetate, trifluoroacetate, adipate, citrate, aspartate, ben/oate. benzcncsulphonatc, bisulfate. bcs\ late, butyrate, camphorsulphonate, difluconae, hemisulfate. heptanoate, formate, fumarate, lactate, maleate.
methanesulfonate, naphthylsulfonate. nicotinale. oxalate, picrate. pivalatc. succinate, tartrate. tirchloracetat, glutamate. p-toluenesulphonate. Indrochloric. hvdrobromic, sulphuric, phosphoric and the like. Representative salts of acidic compounds of the present invention can be prepared by reacting free acid form of the compound with a suitable base, including, but not limited to ammonium, calcium, magnesium, potassium, sodium salts, salts of primary, secondary and tertian amines. substituted amines including naturally occurring ones e.g.. argininc, betaine, caffeine, choline, glucamine, glucosamine, histidine, lysine, morpholine, piperazine, piperidine, purine, triethylamine and the like. Compounds of the present invention that contain a carboxylic acid (-COOH) or alcohol group, their pharmaceutical^ acceptable esters of carboxylic acids such as methyl, ethyl and the like, or acyl derivatives of alcohols such as acetate and the like, can be employed. Compounds of the present invention that comprise basic nitrogen atom may be quaternized with alkyl halides. alkyl sulfates and the like. Such salts permit the preparation of both water soluble and oil soluble compounds of the present invention. It should be recogni/cd that the free base or free acid forms \s ill typically differ from their respective salt forms somewhat in physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free forms for the purpose of the invention.
The "pharmaceutically acceptable solvates" refer to solvates with water (i.e., hydrates) or pharmaceutically acceptable solvents, for example, ethanol and the like.
The invention also encompasses "prodrugs" of the compounds of the present invention which upon in-vivo administration undergo chemical conversion b> metabolic processes before becoming active pharmacological substances. In general such prodrugs will be functional derivatives of a compound of the invention which are readily convertible /;/ vivo into the compound of the invention. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Targeted prodrug design to optimize drug delivery". AAPS I'hariiniSn 2000,2(1), E6.
The invention also encompasses active "metabolites" of the compound of the present invention. An active metabolite is an active derivative of a DPP-IV inhibitor produced when the DPP-1V inhibitor is metabolized.
Various "polymorphs" of a compound of general Formula I forming part of this invention may be prepared by crystallization of a compound of Formula 1 under different conditions, lor example, by using different solvents commonly used or their mixtures for recrystallization; crystallizations at different temperatures; various modes of cooling, ranging from very fast to very slow cooling during crystallizations, heating 01 melting the compound followed by gradual or fast cooling may also obtain polymorphs. The presence of polymorphs may be determined by solid probe NMR spectroscopy, IR spectroscopy, differential scanning calorimetry. powder X-ray diffraction or such other techniques.
The present invention also provides pharmaceutical compositions, comprising compounds of the present invention or their pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof optionally in combination with one or more pharmaceutically acceptable carriers comprising excipients and auxiliaries, flic pharmaceutical compositions may be in any form k noun 111 the art, such as tablets, capsules, powders, syrups, solutions, suspensions and the like. ma> contain tlavourants. sweeteners etc in suitable solid or liquid carriers or diluents, or in suitable sterile media to form injectable solutions or suspensions. Such compositions typically contain active compound optionally in combination with pharmaceutically acceptable carriers, diluents or solvents.
The pharmaceutical compositions of the present invention can be manufactured by the processes well known in the art, for example, by means of conventional mixing, dissolving, dry granulation, wet granulation, dragee-making, levigating, emulsifying, encapsulating, entrapping, lyophilizing processes or spray drying. 1 he compounds or the pharmaceutical compositions comprising such compounds of the present invention may be administered in the form of any pharmaceutical formulation. The pharmaceutical formulation will depend upon the nature ol' the active compound and its route of administration. Any route of administration may be used, for example oral, buccal, pulmonary, topical, parenteral (including subcutaneous, intramuscular and intravenous), transdermal, ocular (ophthalmic), by inhalation, intranasal, transmucosal, implant or rectal administration. Preferably the compounds of the present invention are administered orally. parenterally or topically.
In an embodiment, the amount of the novel compounds having the Formula I according to the present invention to be incorporated into the pharmaceutical compositions of the present invention can vary over a wide range depending on known factors such as. for example, the disorder to be treated, the severity of the disorder, the patient's body weight, the dosage form, the chosen route of administration and the number of administration per day. lypically, the amount of the compound of Formula I in the pharmaceutical compositions of the present invention will range from approximately 0.01 mg to about 5000 mg. In an embodiment, the daih dose of composition comprising the novel compounds having the Formula I is in the range of about 0.01 mg/kg to about 100 mg/kg based on the body weight of the subject in need thereof which may be administered as a single or multiple doses
In an embodiment, the novel compounds having the Formula I according to the present invention are particularly useful for the treatment of discase(s) or disorders) which are particularly acute in nature and which require a short term but mild to moderate treatment, or even some chronic conditions which favorably respond to or are alleviated by the novel compounds having the Formula 1 or compositions comprising them. The compositions comprising the novel compounds having the Formula-I are useful prophylactically or therapeutically depending upon the pathological condition intended to be prevented or treated respectiv ely
The DPP-IV inhibitors of the present invention are useful for the prophylaxis, amelioration and/or treatment of Type 2 diabetes and in the prophylaxis, amelioration and/or treatment of the numerous conditions that often accompany Type 2 diabetes.
The diseases, disorders and conditions mat are related to Type 2 Diabetes and therefore may be treated, controlled in some cases prevented, by treatment with DPP-IV inhibitors include, but not limited to, for example, hyperglycemia and Metabolic Syndrome or 'Syndrome X', including impaired glucose tolerance, insulin resistance, metabolic acidosis or ketosis, disorders of food intake, satiety disorders, obesity, dyslipidemia (including lnpcrlipidemia, hypertriglyceridemia. hypercholesterolemia, low HDL levels, high I.D1. levels), atherosclerosis and its 30 sequele. hypertension associated with metabolic disorders.
Other inflammatory conditions, include but are not limited to, for example, irritable bowel disease(IBS), inflammatory bowel disease including Crohn's disease and ulcerative colitis, pancreatitis, neurodegenerative disease, retinopathy, nephropathy, neuropathy, ovarian hyperandrogenism (polycystic ovarian syndrome) and other disorders where insulin resistance is a component.
Furthermore, the compounds of the present invention may also be useful for the prophylaxis, amelioration and/or treatment of wound healing, tissue ischemia, cataracts, glaucoma, increased cardiovascular risk, growth hormone deficiency neutropia. neuronal disorders, tumor invasion and metastasis, benign prostatic hypertrophv (HIM1). gingivitis, osteoporosis, sperm motility'male contraception, pain, neuropathic pain, rheumatoid pain, osteoarthritis pain, acne, skin disorders (e.g. pigmentation disorders or psoriasis), anxiety, anorexia, epilepsy, male and female sexual dysfunction, major depression disorder, Parkinson's disease, migraine, osteoarthritis, immunosuppression, HIV infection, hematopoiesis, anaemia and other conditions manifested by a variety of metabolic, neurological, anti-inflammatory, and autoimmune disorders including, for example, rheumatoid arthritis, viral, cancer and gastrointestinal disorders that may be prevented or treated by inhibition of DPP-IV.
A further embodiment of the present invention is the use of a compound of Formula 1 for the manufacture of a medicament for the prophylaxis, amelioration and/or treatment of one or more one or more condition(s)/disease(s)/ disorders) mediated by DPP-IV in a subject in need thereof.
Another embodiment of the present invention provides methods for the prophylaxis, amelioration and/or treatment of one or more one or more condition! s)/discasccarbonyl-amino)-acctic acid methyl ester To a solution of ter/-butoxycarbonyl-(2-nitro-rKMi/.\l)-amino|-acetic acid methyl ester (845 mg. 2.61 mmol) in MeOH (15 mL), was addedl0% Pd/C" (84.5 mg. 10% b\ weight) under nitrogen atmosphere. The reaction mixture was evacuated and then charged with hydrogen gas and hydrogenated using hydrogen balloon for 3 h. After completion of the reaction as confirmed by TLC. the reaction mixture was evacuated and brought under nitrogen atmosphere and then filtered through celite bed, washed with MeOH (10 mL). The filtrate was concentrated in vacuo to afford [(2-amino-benzyl)-/i'r/-butoxycarbonyl-amino]-acetic acid methyl ester (660 mg, 86%) as a yellow oil which was used as such without any further purification for the next step. ESIMS (m/z): 295.4 (M+l)
Step 4: Preparation of 2-oxo-l^r33-tetrahydro-benzo|^||l,4|diazepine-4-carboxylic acid tert-butyl ester
To a solution of [(2-amino-benzyl)-/£V7-buu>x\carbon\l-amiru)|-aeetic acid mctlnl ester (640 mg. 2.18 mmol) in toluene (30 mL), was added HOHl (177 mg, 1.31 mmol). The reaction mixture was heated at 100 °C for 24 h. After the completion of reaction, as confirmed by TLC', the solvent was removed in vacuo to afford the crude compound, which was purified by column chromatography (silica gel, 7:13 EtOAc: Pet. Ether) to afford pure 2-o\o-l,2.3,5-tctrahydro-benzo|c|| l,4|diazepine-4-carboxylic acid ter/-butyl ester (350 mg, 61%) as a solid
ESIMS (m/z): 263.3 (M+l)
Step 5: Preparation of trifluoroacetic acid salt of l,3,44^tetrahydro-bcnzo[£](l,4|diazepin-2-one To a solution of 2-oxo-l,2,3,5-tetrahydro-ben/.o|^|[l,4|diazepine-4-carboxylic acid /t?r/-but>l ester (300 mg, 1.14 mmol) in dry DCM (10 mL) was added trifluoroacetic acid (3.42 mL. 3mL/mmol). The reaction mixture was stirred at r.t. for 2 h. Alter completion of the reaction as confirmed b\ II C . excess of trifluoroacetic acid and DCM were removed under vacuo to afford trifluoroacetic acid ->all of l,3,4,5-tetrahydro-benzo[e][l,4]diazepin-2-one (2l>> mg. 93 %) as a gummy solid. I he crude compound was used as such without further purification for the next step.
ESIMS (m/z): 185.3 (M+Na), 163.3 (Mi 1)
Step 6: Preparation of |(/?)-3^xo-3-(2-oxo-l^,3,5-tetrahydro-benzo[e][l,4]diazepin-4-yl)-l-(2,4,5-trifliioro-beiizyl)-propyl]- carbamic acid /erf-butyl ester
To a solution of (/?)-3-[(/er/-butoxycarbonyl)aminol-4-(2.4,5-tritluorophenyl)butanoic acid (90 mg, 0.26 mmol) in dry DCM ( 10 mL), was added HOB! (47 mg, 0.34 mmol), EDC (67 mg, 0.34 mmol) and DIPEA (0.24 mL, 1.35 mmol). The reaction mixture was stirred at r.t. for 10 min. A solution of l,3,4,5-Tetrahydro-benzo[e][l,4]diazepin-2-onc trifluoroacetate (74 mg, 0.26 mmol) in dr> IX'M (5 mL) was added and the reaction mixture was stirred at r.t. overnight under nitrogen atmosphere. After
the completion of the reaction, as confirmed b\ TLC. the crude product was extracted with IX'M (10 mL) and washed sequentially with 10% IK I solution (10 ml.) and saturated sodium bicarbonate solution (10 mL). The organic layer was separated, washed with water, dried over Na^SOj and concentrated in vacuo to afford the crude compound, which was purified by column chromatography (silica gel, 2:3 EtOAc: Pet. Ether) to afford r(/0-3-Oxo-3-(2-oxo-1.2,3,5-tetrah\dro-benzo[e][l,4]diazepin^-yl)-l-(2,4,5-trifluoro-benzyl)-propyl|- carbamic acid /e/7-butyl ester (80 mg, 62%) as a white solid.
ESIMS (m/z): 500.2 (M+Na), 478.2 (M i I i
Step 7: Preparation of tnfluroacetic acid salt of (/?)-4-|3-Amino-4-(2,4,5-trifluorophcnyl)-bntyryfJ-l,3,4,5-tetrahydro-benzo[e][l,4]diazepin-2-one
To a solution of [(7?)-3-oxo-3-(2-o.\o-1.2. i.5-tetrahydro-bcn/o|t'][ l,4|dia/.epin-4-yl)-l-(2.4.5-trifluoro-benzyl)-propyl]- carbamic acid teri-bu\\\ ester (40 mg, 0.08 mmol) in DCM (5 ml.), was added trifluoroacetic acid (0.24 mL, 3mL/mmol). I he reaction mixture was stirred at r.t. for 1 h. Alter completion of the reaction, as confimed b> II.C. excess of trifluoroacetic acid and DCM were evaporated in vacuo to afford a gummy solid which was solidified from hexane to afford tnfluroacetic acid salt of 4-[(/?)-3-amino-4-(2,4,5-trifluorophenyl)-but>'ryl|-l,3,4,5-tetrahydro-benzo[4'|f 1,41 diazepin-2-one (30 mg, 73%) as a solid.
*H NMR (400 MHz, MeOD): 8 2.62-2.70 (m. Ill), 2.75-2.87 (m. Ill), 2.97-3.03 (m, 211). 3.76-3.86 (two m, 1H), 4.17 (s, 1H), 4.50 (s. 1H). 4.62 (s. III). 4 66 (s. III). 7.05-7.15 (m. 211). 7.14-7 25 (m. 2H), 7.27-7.30 (m, 211) ESIMS (m/z): 400.3 (M+Na), 378.2 (Mi 1)
Example 2: Preparation of trifluoroacetic acid salt of (/f)-4-(3-amino-4-{2,4,5-trifluoiX)phenyl)butanoyl)-73^ifluoro-4,5-dihydro-lH-benzo[^||l,41diazepin-2(3H)--one
(Formula Removed)
Step 1: Preparation of 2-amino-4,5-diflouro-phcnyl-mcthanol
To a solution of lithium aluminium hydride (2.1 ^ g. 5" 76 mmol) in dry THI- (35 ml.) at 0 '( . was slowly added a solution of 2-amino-4,5-diflouro benzoic acid (5 g. 28.88 mmol) dissolved in dr> l"Hr (35 mL). The reaction was stirred at r.t. for 2 h. After completion of the reaction as confirmed b\ TLC, water (2.1 mL), 10 % NaOH (2.1 ml.) and water (3x 2.1 mL) were added sequentially to the reaction mixture at 0 °C. The resultant sluny was stirred at r.t.for 20 min. The slurry was filtered from celite bed and washed with EtOAc (2x20 ml.). I he filtrate was dried over Na2S04 and the solvents
were removed in vacuo to afford the crude compound as a solid. The crude compound, (2-amino-l. 5-
diflouro-phenyl)-methanol (4.50 g, 98%) was used as such without any further purification for the
next step.
ESIMS(m/z): 158.1 (M-l)
Step 2: Preparation of tert-butyl 4,5-difluoro-2-(hydroxymethyl)phcnylcarbamatc
To a solution of (2-amino-4,5-difluoro-phen\ I (-methanol (4.50 g, 28.30 mmol) in 1, 4-dioxane and
water (1:1, 50 mL) was added NaHC03 (4.7 g. 56.60 mmol) at 0 °C. To the resulting solution, was
added di-/er/-butyl dicarbonate (9.7 mL, 42.45 mmol). The reaction was stirred at r.t. for 10 h. After
completion of the reaction, as confirmed by II X\ 1. 4-dioxane was removed in vacuo and crude
compound was extracted with EtOAc (3\50 ml.). The combined organic layers were dried over
Na2SC>4 and the solvents were removed in vacuo lo afford the crude compound, which was purified by
column chromatography (silica gel, 1:9 1-tOAc: Pel. I.ther) lo afford pure (4. 5-dilluon>-2-
hydroxymetyhl-phenyl)-carbamic acid /cr/-but\ I ester (5.8ft it. 80%) as a thick gel.
ESIMS(nVz): 258.3 (M-l)
Step 3: Preparation of ferf-butyl 2-(chloromethyl)-4,5-difluorophenylcarbamate
To a solution of (4, 5-difluoro-2-hydroxymetyhl-phenyl)-carbamic acid te/7-butyl ester (5.86 g, 22.62
mmol) in dry DCM (50 mL) was added thion\ I chloride (3.28 mL, 45.64 mmol) at 0 °C. The reaction
was stirred at r.t. for 2 h under argon. After completion of the reaction as confirmed by TLC. water
(30 mL) was added to the reaction mixture and crude compound was extracted with DCM (3x30 mL).
The combined organic layers were dried over Na^SOi and the solvents were removed in vacuo to
afford the crude (2-chloromethyl-4, 5-difluoro-phenyl)-carbamic acid /erf-butyl ester (5.65 g, 90%)
which was used as such without any further purification for the next step.
ESIMS(nVz): 279.3 (M+l)
Step 4: Preparation of (2-/«rf-butoxycarDonylamino-4, 5 difluoro-benzylaniino)-acetic acid
methyl ester
To a solution of (2-chloromethyl-4, 5-difluoro-phenyl)-carbamic acid tert-buiy\ ester) (5.65 g. 20.3
mmol) in DMF (50 mL) was added DIPEA (10.6 ml.. 60.9 mmol). The resulting solution was stirred
for 5 min, followed by dropwise addition of solution of glycine methyl ester hydrochloride in DM1- at
0 °C. The reaction mixture was stirred overnight at 50 °C. After completion of the reaction as
confirmed by TLC, water (60 mL) was added to the reaction mixture and crude compound was
extracted with EtOAc (3x50 mL). The combined organic layers were dried over Na2SO». The solvents
were removed in vacuo to afford the crude compound which was purified by column chromatography
(silica gel: 1.5: 8.5 EtOAc: Pet. ether) to obtain pure (2-^r/-butoxycarbonylamino-4, 5 difluoro-
benzylamino)-acetic acid methyl ester (5.37 g. X0%) as thick gel.
ESlMS(m/z): 331.4(M+1)
Step 5: Preparation of benzyloxy carbonyl-(2-fc/7-butoxycarbonylaniino-4, 5-difluoro-bcnzyl)-
amino]-acetic acid methyl ester
To a solution of (2-terf-butoxycarbonylamino-4. 5 difluoro-benzylamino)-acetic acid methyl ester (5.37 g, 16.28 mmol) in 1,4-dioxane: water (1:1. 50 ml.) was added NaHCO, (2.73 g. 32.56 mmol) at 0 °C, followed by the addition of benzyl chlorofromate (4.09 ml.. 24.42 mmol). The reaction was stirred at r.t. for 10 h. After completion of the reaction as confirmed In 111'. '. 1-dioxanc was removed in vacuo and crude compound was extracted v\ith KtOAc (3x50 ml.). 1 he combined organic-layers were dried over Na2S04 and the solvents were removed in vacuo to afford the crude compound, which was purified by column chromatography (silica gel. 1.5: 8.5 KtOAc: Pet. ether) to afford pure [benzyloxy carbonyl-(2-ter/-butoxycarbonylamino-4. 5-defluoro-benzyl)-amino|-acctie acid metlnl ester (5.28 g, 70%) as thick gel. ESIMS(nVz): 463.4(M-1)
STEP 6: Preparation of benzyloxy carbonyl-(2-ferf-butoxycarbonylamino-4, 5-difiuoro-benzyl)-amino]-acetk acid
To a solution of benzyloxy carbonyl-(2-/t'/7-butox\carbonylamino-4, 5-difluoro-benzyl)-amino|-acetic acid methyl ester (5.28 g, 11.39 mmol) in THF (30 mL) was added a solution of lithium hydroxide (2.87 g, 68.34 mmol) in water (10 ml.). I he reaction was stirred at r.t. lor 10 h. A tier completion of the reaction as confirmed b> II.( . the reaction mixture was acidified to pi 1 4 b\ adding 10% HC1 and crude compound was extracted with KtOAc (3x50 mL). The combined organic lavers were dried over Na2SC>4 and the solvents were remtned in vacuo to afford the crude compound [benzyloxy carbonyl-(2-/e/7-butoxycarbon\lamino-4. 5-difluoro-ben/yl)-amino|-acetic acid (4.76 g. 93%) as a solid, which was used as such w ithout an\ further purification for the next step. ESIMS(m/z):448.8(M-l)
Step 7: Preparation of 2-[(benzyloxycarbonyl-carboxymethyl-aniino)-methyl]-4^-difluoro-phenyl-ammonium trifluoroacetate
To a solution of {benzyloxy carbonyl-(2-/(T/-butoxycarbonylamino-4,5-difluoro-benzyl)-amino|-acetic acid}(4.76 g, 10.56 mmol) in dry FX'M (40 ml.) was added trifluoroacetic acid (14.35 ml.. 3 mlVmmol) at 0 °C. The reaction mixture was stirred at r i lor 2 h under argon After completion of the reaction as confirmed by TLC, the reaction mixture was evaporated in vacuo to remove excess of solvent and trifluoroacetic acid. The remaining solvents were removed under vacuo and 2-[(benzyloxycarbonyl-carboxymcthyl-amino)-methylj-4.5-difluoro-phenyl-ammonium trifluoroacetate (4.75 g, 97%) was obtained as thick brown jelK which was used as such without any further purification for the next step. ESIMS(m/z): 351.2 (M+l)
Step 8: Preparation of benzyl 7^-difluoro-2-oxo-23-dihydro-l/f-benzo[«|[l,4|diazepine-4(5H)-carboxylate
To a solution of 2-[(benzyloxycarbon\l-carbox\methyl-amino)-methylJ-4,5-difluoro-phenyl-ammonium trifluoroacetate (4.75 g, 10.25 mmol) in DCM (40 mL) was added EDC (2.55 g. 13.32mmol) and HOBT (1.79g, 13.32 mmol) at 0 °(\ The resulting solution was stirred for 5 min and
DIPEA (5.35 mL, 30.75 mmol) was added to it. The reaction mixture was stirred overnight at r.t.
under argon. After completion of the reaction as conllnned by PLC. water (60 mL) was added to the
reaction mixture and crude compound was extracted with DCM (3x50 mL). The combined organic
layers were dried over Na2SC>4 and the solvents were removed in vacuo to afford the crude compound
which was purified by column chromatography (silica gel: 2: 8 litOAc: Pet. ether) to afford pure
benzyl 7,8-difluoro-2-oxo-2,3-dihydro-lH-ben/o|i'|| 1.4|diazepine-4(5H)-carboxylate (2.04 g. 60%)
as thick gel.
ESIMS(m/z):331.2(M-l)
Step 9: Preparation of 7, 8-difluoro-l, 3, 4, 5-tetrahydro-ben/o|?| [1, 4) dia/.epin-2-onc
To a solution of (7, 8-difluoro-2-oxo-l, 2. 3. 5-tetrahydro-benzo[e] 11, 4] diay.epine-4-carboxyiic acid
phenyl ester) (2.04 g, 6.15 mmol) in MeOH (30 ml.), was added 10% Pd/C (200 mg. 20% WH) under
argon. The reaction was stirred at r.t. for 5 h under hydrogen atmosphere. After completion of" the
reaction as confirmed by TLC, the reaction mixture was filtered through celite bed and washings were
given with EtOAc and MeOH. The solvents were removed in vacuo to afford the crude compound
which was purified by column chromatography (silica gel: 1:9 MeOH: CHCI3) to afford pure 7. 8-
difluoro-1, 3, 4, 5-tetrahydro-benzo[e] [1, 4| diazepin-2-one as off white solid (0.98 g, 80%).
ESIMS(nVz): 199.7 (M+l)
Step 10: Preparation of (R)-tert-butyl 4-(7,8-difluoro-2-oxo-2r5-dihydro-lH-
benzo[e][l,4Jdiazepin-4{5W)-y0-4-oxo-l-(2,4,5-trifluorophcnyl)butan-2-ylcarbaniatc
To a solution of (/?)-3-[(/e/,/-butoxycarbonyl)amino|-l-(2,4,5-lrifliiorophcnyl)bulaiioic acid (1.61 g.
4.92 mmol) in dry DCM (15 mL) was added l.DC (1.037 g. 5.412 mmol) and HOB I" (0.731 g. 5.412
mmol) at 0 °C. The resulting solution was stirred for 5 min and DIPEA (0.94 mL, 5.412 mmol) was
added to it, followed by the addition of 7. 8-difluoro-l. 3. 4. 5-tetrahydro-benzo[fJ 11, 4] diazepin-2-
one (0.97 g, 4.92 mmol). The reaction mixture was stirred for 5 h at r.t. under argon. After completion
of the reaction as confirmed by TLC, water (30 ml.) was added to the reaction mixture and crude
compound was extracted with DCM (3x30 ml). The combined organic layers were dried over
Na2S04 and the solvents were removed in vacuo to afford the crude compound which was purified by
column chromatography (silica gel: 0.7: 99.3 MeOH: CHCI3) to afford pure |3-(7.8-difluoro-2-oxo-
l,2,3,5-tetrahydro-benzo[e][l,4Jdiazepine-4-\l)-3-oxo-l-(2.4,5-trifluorzyl)-propyl]-carbamic acid
/ert-butylester (0.682 g. 27%).
'H NMR (400 MHz, CDCI.,): 8 1.35 and 1.38 (two rotameric singlets. 911). 2.54-2 65 (m. 211). 2.8l>-
2.92 (m, 2H), 4.12 (m, 1H), 4.17-4.20 (m. 111). 4.44-4.62 (m. 311). 5.38-5.43 (m. 111). 6.84-6.l)() im.
2H), 6.98-7.21 (m, 2H), 8.11 and 8.31 (two bs. III)
ESIMS(nVz): 514.4(M+l)
Step 11: Preparation of trifluoroacetic acid salt of (^?)-4-(3-aniino-4-(2,4,5-
trifluorophenyl)butanoyl)-73-difluoro-4^-dihydro- lH-benxo[e j [ 1,4]diazepin-2(3H)-one
To a solution of [3-{7,8-difluoro-2-oxo-l,2.V5-tetrahvdro-ben/o|^|| l,4|dia/epinc-4-vl)-3-o\o-l-(2,4,5-trifluorzyl)-propyl]-carbamic acid /cW-butylester. (0.682 g. 1.32 mmol) in drv DCM (S ml.) was added trifluoroacetic acid (3.96 mL, 3ml./mmol) at 0 "(.'. The reaction mixture was stirred at r.t. for 2 h under argon. After completion of the reaction as confirmed by TLC, the reaction mixture was evaporated under vacuo to remove excess of solvent and trifluoroacetic acid. The remaining solvents were removed under vacuo and the crude compound was triturated with diethylethcr (5 mL) resulting in trifluoroacetic acid salt of (^?)-4-(3-amino-4-(2.4.5-trifluorophenyl)butanoyl)-7,8-difluoro-4.5-dihydro-lH-benzo[e][l,4]diazepin-2(3H)Hine (0.63 g. 90%) as a white solid.
*H NMR (400 MHz, CDC13): 5 2.65-2.88 (m. 2H). 2.97-3.08 (m, 211), 3.81-3.92 (m. 1H), 4.22 (s. 1 H). 4.45^.64 (m, 3H), 7.01-7.33 (m, 411)
ESIMS(m/z): 415.6(M+1)
Example 3: Preparation of trifluroacetie acid salt of 4-|(/?)-3-amino-4-(2, 4, 5-trifluorophvnvl)-
butyryl]-l-methyl-l^,4^-tetrahydro-ben«)(^| 11,4|diazcpin-2-onc
(Formula Removed)
Step 1: Preparation of l-methyl-2-oxo-U3,Metrahydro-ben,«)lelll,41diaxepinc-4-carboxylic acid tert-buryl ester
Sodium hydride (18 mg, 0.45 mmol, 60% suspension in mineral oil) was washed with hexane (2*2 mL) in a flame dried round bottomed llask uiulei nilrouon atmosphere, lo the resulting free lloatmg powder, was added a solution of 2-oxo-1.2.3.5-tetrah>dro-bcn/o|e|| l,4|diazcpinc-4-carboxylic acid tert-buty\ ester ( 80 mg, 0.30 mmol) in dry DMh (3 ml.) at 0 °C. 1 he reaction mixture was stirred at this temperature for 1 h. lodomethane (0.019 ml., 0.30 mmol) was added at 0 °C and the reaction mixture was allowed to come to r.t. and stirred overnight. After the completion of the reaction as confirmed by TLC, reaction mixture was cooled to 0 °C and saturated solution of ammonium chloride (2 mL) was added dropwise to the reaction mixture, fhe crude product was extracted with ethyl acetate (2x10 mL). The combined organic layers were dried over NaiSOi and the solvents were removed in vacuo to afford l-methyl-2-oxo-l.2,3,5-tetrahydro-bcnzo[e][l,4]diazepine-4-carboxylic acid tert-buty\ ester (75 mg, 89%) as a viscous oil which was used as such without an> further purification for the next step. ESIMS (m/z): 278.3 (M+2). 277.3 (M-t 1)
Step 2: Preparation of trifluoroacetic acid salt of l-methyl-l,3,4,5-U:trahydrobcn/.o |c| [l,4|diazepin-2-one
To a solution of l-methyl-2-oxo-l,2,3,5-tetrahydro-benzo[eJ| 1.4|diazcpine-4-carboxylic acid tert-butyl ester (70 mg, 0.25 mmol) in dry DCM (10 ml.), was added trifluoroacetic acid ( 0.75 int., 3mL/mmoI). The mixture was stirred at r.t for lh. Alter the completion of the reaction as confirmed by TLC, excess of TFA and DCM were evaporated in vacuo lo afford trilluoroacclic acid salt of I-methyl-l,3,4,5-tetrahydrobenzo [e] [l,4Jdia/epin-2-one
(70 mg, 95%) as a gummy solid which was used as such for next coupling.
Step 3: Preparation of 2 [3-(l-methyl-2-oxo-l^r3^-tetrahydro-benzo[c||l,4|diazcpin-4->l>-3-
oxo-(l?)-l-(2,4,5-trifluoro-benzyl) -propylJ-carbamic acid tert-butyl ester
To a solution of (J?)-3-[(/e/7-butoxycarbonyl)amino|- l-(2,4.5-trifluorophenyl)butanoic acid (80 ing.
0.24 mmol) in dry DCM (10 mL), was added HOBT (42 mg, 0.31 mmol), EDC (59 mg, 0.31 mmol)
and D1PEA (0.20 mL, 1.2 mmol). The reaction mixture was stirred at r.t. for 10 min. A solution of 1-
methyl-l,3,4,5-tetrahydrobenzo (e) [l,4]dia/.epin-2-one trifluoroaccte (70 mg, 0.24 mmol) in dry
DCM (5 mL) was added and the reaction mixture was stirred at r.t. overnight. After the completion of
the reaction, as confirmed by TLC. the crude product was extracted with DCM (10 mL) and washed
sequentially with 10% HCI solution (10 ml ) and saturated solution of sodium bicarbonate (ll) ml )
The organic layer was separated and washed with brine and dried over Na-SO,. The solvents were
removed in vacuo to afford the crude compound, which was purified by column chromatography
(silica gel, 3:7 EtOAc: Pet. Ether) to atl'ord 2 |3-(l-mcthyl-2-oxo-1.2.3.5-tetralndro-
benz»[^][l,4]diazepin-4-yl)-3-oxo-(^?)-l-(2,4.5-trinuoro-ben/yl) -propylj-carbamic acid icri-by\[\ester (95 mg, 78%) as viscous oil.
ESIMS(m/z): 492.3 (M+l)
Step 4: Preparation of trifluroacetic acid salt of 4-l(i?)-3-amino-4-(2,4,5-trifluorophenyl)-
butyryl]-l-methyH^,4^-tetrahydro-benzo|«?)|l,4]diazcpin-2-onc
To a solution of 2 [3-(l-methyl-2oxo-l,2.3.5-tetrahydro-bcnzo|eJ| l,4Jdiazepin-4-yl)-3-oxo-(/?)-l-(2,4,5-trifluoro-benzyl) -propylj-carbamic acid /cry-butyl ester (90 mg, 0.18 mmol) in dry DCM ( 5 mL), was added trifluoroacetic acid (0.55 ml . 3 ml mmol). I he reaction mixture was stirred at it for lh. After completion of the reaction, as confirmed b\ II.C. excess of MA and IX VI were evaporated in vacuo to afford a gumm\ solid which was crystallised from hexane to aflord trifluroacetic acid salt of 4-[(/?)-3-amino-l-(2.-1.5-trifluorophenyl)-bu1>r>l|-I-methyl-l.3,4,5-tetrahydro-benzo[e][l,4]diazepin-2-one (70 mg. 76% ) as a white solid
'H NMR (400 MHz, MeOD): 8 2.72-2.89 (m. 211). 2.09-3.09 (m. 211), 3.40 (s, 3H), 3.88-3.92 (in. 2H), 4.05-4.15 (m, 1H), 4.50-4.58 (m, 2H), 7.20-7.25 (m. 111). 7.28-7.53 (m, 511) ESIMS (m/z): 415.4 (M+Na), 391.1 (M-H )
Example 4: Preparation of trifluroacetic acid salt of {4-|(.tf)-3-amino-4-(2,4,5-trifluoro-phenyl)-butyryl]-2-oxo-2,3,4,5-tetrab.ydro-benzolc|| 1.4|diazepin-l-yl}-acctic acid
(Formula Removed)
Step 1: Preparation of l-ethoiycarbonylraethyl-l-oxo-l^^^-tetrahydro-benzolc'Ul^ldiazepine-
4-carboxylic acid fcrr-butyl ester
Sodium hydride (152.6 mg, 3.81 mmol. 60u/<> suspension in mineral oil) was washed w ith hcxune 1 2 3
mL) in a flame dried round bottomed flask under nitrogen atmosphere. To the resulting free floating
powder, was added a solution of 2-oxo-1.2.3.:>-tetrah\dro-ben/.o|c|| l,4Jdiaxcpine-4-carboxylic acid
terr-butyl ester (500 mg, 1.90 mmol) in dry DM1- (10 ml.) at 0 °C\ The reaction mixture was stirred at
this temperature for lh. Ethyl bromoacetatc (0.21 mL. 1.9 mmol) was added at 0 °C and the reaction
mixture was allowed to come to r.t. and stirred overnight. After the completion of the reaction as
confirmed by TLC, reaction mixture was cooled to 0 °t" and saturated solution of ammonium chloride
(3 mL) was added dropwise to the reaction mixture. The crude product was extracted with
ethylacetate (3><10 mL). The combined organic layers were dried over NaiSC^ and solvents were
removed in vacuo to afford l-cthox\carbonylmethyl-2-oxo-1.2,3,5-tetrahjdro-
benzo[e][l,4]diazepine-4-carboxylic acid /i'H-butyl ester (480 mg, 72%) as a viscous oil which was used as such without any further purification for the next step.
ESIMS(m/z): 349.1 (M+l)
Step 2: Preparation of trifluoroacetic acid salt of (2-oxo-2,3,4,5-tetrahydro-
benzo[e][l,4]diazepin-l-yl)-acetic acid ethyl ester
To a solution of l-ethoxycarbonylmetli\l-2-oxo-1.2,3..S-tclrahydro-ben/o|c|| l,4jdia/cpinc-4-carboxylic acid tert-butyl ester (125 mg, 0.36 mmol) in dry IX'M (5 mL) was added trifluoroacetic acid (1.08 mL, 3 mL/mmol). The reaction mixture was stirred at r.t. for one hour. After completion of the reaction, as confirmed by TLC, excess of trifluoroacetic acid and DCM were evaporated in vacuo to afford trifluoroacetic acid salt of (2-oxo-2.3.4.5-tetrahydro-benzole][l,4]diazepin-l-yl)-acetic acid ethyl ester (119 mg, 92 %) as a gummy solid which was used as such without any further purification for die next step.
Step 3: Preparation of {4-|3-/er/-butoxyearbonylamino-4-(2,4,5-trifluoro-phcnyl)-but>rvl|-2-oxo-2,3,4,5-tetrahydro-benzo[e][l,4]diay.cpin-l-ylJ-acctic acid methyl ester
To a solution of (/?)-3-[(fcrf-butoxycarbonyl)amino|-4-{2,4,5-trifiuorophenyl)butanoic acid (100 mg, 0.30 mmol) was dissolved in dry DCM (10 ml), was added MC)BT(53 mg, 0.39 mmol), EDC(75 mg, 0.39 mmol) and DIPEA (0.26 mL, 1.49 mmol) . I he reaction mixture was stirred at r.t. for 10 mm. A solution of (2-oxo-2,3,4,5-tetrahydro-bcn/o[f || 1.4|dia/.epin-l-\l)-acctic acid cth\ I ester trifluoroacetate (119 mg, 0.33 mmol) in IX'M (5 mL) was added and the reaction mixture was stirred at r.t. overnight under nitrogen atmosphere. After the completion of the reaction, as confirmed by
TLC, the crude product was extracted with DCM (10 ml.) and washed sequentially with 10% MCI (10 mL) and saturated solution of sodium bicarbonate (10 ml,). The organic layer was separated, washed with water, dried over Na2SC>4 and concentrated in vacuo to afford the crude compound, which was purified by column chromatography (silica gel. 1:1 LtOAe:Pet Ether) to afford |4-|3-/e/7-butoxycarbonylamino-4-(2,4,5-trifluoro-phen> I )-butyr\ I |-2-oxo-2,3.4,5-tetrahydro-benzo[e][l,4]diazepin-l-yl}-acetic acid methyl ester (103 mg, 61 %) as a viscous oil. ESIMS (m/z): 586.1 (M+Na), 564.0 (M-t 1)
Step 4: Preparation of {4-[3-/«/f-butoxycurbonylamino-4-<2,4^-trifluoro-phenyl)-butyryl]-2-oxo-2,3,4,5-tetrahydro-benzo[el[l,4]diazepin-l-yl}-acetic acid
To a stirred solution of {4-[3-te^butoxycarrR>nylamino-4-(2,4,5-trifluoro-phenyl)-butyryl|-2-oxo-2,3.4,5-tetrahydro-benzo[e]| 1,4 |diazepin-l-> 11-acetic acid methsl ester (90 mg. 0.16 inmol) in I III (2 mL) and water (2 mL), was added lithium h\droxidc (40 mg. 0.95 mmol). I'he reaction mixture was stirred at r.t. overnight. After the completion of the reaction as confirmed by TLC. the reaction mixture was cooled to 0° C and neutralized with 10 % MCI solution and the pll was adjusted to 1. The crude compound was extracted with ethylacetate (10 ml.). 1 he organic layer was washed with water, dried over Na2SC>4. The solvents were removed in vacuo to afford |4-|3-te/7-buto.\ycarbon\ lammo-4-(2,4,5-trifluoro-phenyl)-butyryl]-2-oxo-2,3.4.5-ietrah>dro-benzo|e|ri,4Jdiazepin-l-yl}-acetic acid (70 mg, 82 %) as a viscous oil.
ESIMS (m/z): 558.3 (M+Na), 536.4 (M+l ). 534.3 (M-l)
Step 5: Preparation of trifluroacetic acid salt of {4-|(/?)-3-amino-4-(2,4,5-trifluoro-phenyl)-butyryl]-2-oxo-2^,4^-tetrahydro-benzo[e||l,4|diazepin-l-yl}-acctic acid
To a solution of {4-[3-te/7-butoxycarbon\ lamino-4-(2.4.5-trilluoro-phcnyl)-butyryl|-2-oxo-2. *.4,v tetrahydro-benzo[e][l,4]diazcpin-l-yl}-acetic acid (60 mg. 0.1 1 mmol) in IX'M (3 ml.), was added trifluoroacetic acid (0.33 mL, 3 mL/mmol). I he reaction mixture was stirred at r.t. for I h Alter completion of the reaction, as confirmed b\ IIX'. excess of trifluoroacetic acid and DCM were evaporated in vacuo to afford a gummy solid which was erystalised from hexane to afford trifluroacetic acid salt of {4-[(fl)-3-amino-4-(2.4.5-trifluoro-phenyl)-butyrYl|-2-oxo-2.K-l.>-tetrahydro-benzo[e][l,4]diazepin-l-yl}-acetic acid (50 mg. 82 %) as a solid.
'H NMR (400 MHz, MeOD): S 2.64-2.90 (m. 2M). 2.98-3.12 (m, 2H), 3.80-3.88 (m, 1H), 3.95 (s, 1H), 4.05-4.17 (m, 1H), 4.61-4.64 (m, 2H), 4.73-4.81 (m. 2H), 7.20-7.40 (m, 6M) ESIMS (m/z): 458.4 (M+Na), 436.3 (M4 1)
Example 5: Preparation of trifluroacetic acid salt of 2-{4-[(X)-3-amino-4-(2,4^-trifluoro-phenyl)-butyryl]-2-oxo-2r3,4^-tetrahydro-ben/.o|(f||l,4)diay.cpin-l-yl}-A/-cyclopropyl-acctamidi'
(Formula Removed)
Step 1: Preparation of [3-0-cycIopropylcarbamoylmethyl-2-oxo-l,2,3,5-tetrahydro-benzo[«][l,4]diazepin-4-yl)-3-oxo-l-(2,4,5-trifluoro-benzyl)-propyl|-carbamic acid tert-butyl ester
This compound was obtained by coupling of |4-|3-/<77-butoxyearbonylamino-4-(2,4,5-trilluoro-
phenyl)-butyryl]-2-oxo-2,3,4,5-tetrahydro-ben/o[i'J[ 1.4 |diazepin-1 -yl} -acetic acid and
cyclopropylamine using HOBT, EDC, DIPEA following the above mentioned procedures.
ESIMS (m/z): 597.5 (M+Na), 575.4 (M+ 1)
Step 2: Preparation of trifluroacetic acid salt of 2-{4-[(/?)-3-amino-4-(2,4^>-trifluoro-phenyl)-butyryi]-2-oio-2^,4^-tetrahydro-benzo|f)(l,4)diay^pin-l-yl}-A'-cyclopropyl-acetamide To a solution of [3-{l-cyclopropylcarbanio\ Imelln l-2-o\o -1.2.3.5-tctrahydro-bcn/o|<'|| I. l|dia/cpin-4-yl)-3-oxo-l-(2,4,5-trifluoro-bcnzyl)-prop\l|-carbamic acid /t'/7-butyl ester (40 mg, 0.07 miiml) in DCM (2 mL), was added trifluoroacetic acid (0.2 ml.. 3 mL/mmol). The reaction mixture was stirred at r.t. for 1 h. After the completion of the reaction as confirmed by TI.C, excess of trifluoroacetic acid and DCM were removed in vacuo to afford a gumim solid which was crystallised from hexane to afford trifluroacetic acid salt of 2-{4-[(/?)-3-amino-4-(2.4.5-trifluoro-phenyl)-butyry l]-2-oxo-2.3.4.5-tetrahydro-benzo[e][l,4]diazepin-l-yl}-N-cyclopropyl-acetamidc (32 mg, 78 %).
'H NMR (400 MHz, MeOD): S 0.46-0.47 (in, 211). 0.68-0.70 (m. 2H), 2.55-2.65 (m, 1H), 2.65-3.07 (m, 2H), 3.01-3.09 (m, 2H), 3.72-3.88 (m. III). 3.95 (s. III). 4.09-4.21 (m, III), 4.45-4.54 (m. 2H). 4.70-4.79 (m, 2H), 7.20-7.50 (m, 6H) ESIMS (m/z): 497.3 (M+Na), 475.1 (M+1)
Example 6: Preparation of trifluoroacetic acid salt of (^)-3-amino-l-(9-flu()n)-4H-benzo[/][l^,4]triazolo[4r3-a)|l,4]diazepin-5(67/)-yl>-4-(2,4^-trifluorophcnyl)butan-l-onc
(Formula Removed)
Step 1: Preparation of (4-fluoro-2-nitro-benzylamino)-acetic acid methyl ester
To a solution of l-bromoethyl-4-fluoro-2-nitrobcn/cne (5.00 g, 21.36 mmol) in DMF (50 ml.) under argon atmosphere, was added DIPEA (11.16 ml.. 64.10 mmol) dropwise at 0 °C. The resulting solution was stirred for 5 min, followed by dropwise addition of solution of glycine methyl ester
hydrochloride (3.48 g, 27.77 mmol) in DMI (5 mL) at 0 °C. The reaction mixture was stirred at r.t.
for 8 h. After completion of the reaction as confirmed b\ NX', water (100 ml.) was added and the
crude compound was extracted with EtOAc (3x200 ml.). The combined organic layers were washed
with brine and dried over Na2SO.i. The solvents were removed in vacuo to afford crude compound
which was purified by column chromatographv |silica gel, 2:8 EtOAc: Pet. ether] to afford 4-fluoro-2-
nitro-benzylamino-acetic acid methyl ester as a dark yellow viscous liquid (4.65 g. 90%).
ESIMS (m/z): 242.8(M+1)
Step 2: Preparation of [ter/-butoxycarbonyM4-nuoro-2-nitro-bcn/.yl)-amino|-acctic acid methyl
ester
To a solution of 4-fluoro-2-nitro-benzylamino-acetic acid methyl ester (4.65 g. 19.21 mmol) in lH'M
(30 mL) under argon atmosphere, was added di-icri-buty\ dicarbonate (4.41 ml.. 19.21 mmol)
dropwise at r.t. The reaction mixture was stirred at r.t.. overnight. After completion of the reaction as
confirmed by TLC, water (30 mL) was added and the mixture was extracted with DCM (3x100 mL).
The combined organic layers were dried over Na^SO, and concentrated in vacuo. The crude
compound was purified by column chromatography [silica gel, 2:8 EtOAc: Pet. ether] to afford pure
/er/-butoxycarbonyl-(4-fluoro-2-nitro-benz>i)-amino-acctic acid methyl ester as a dark yellow liquid
(5.91 g,90%).
ESIMS (m/z): 366.1 (M+Na), 343.2 (M-f 1)
Step 3: Preparation of 2-(^r/-butoxycarbonyl(4-fluoro-2-nitrobeny.yl)amino)-acetic acid
To a solution of crude f/e/7-butoxycarbon\ l-( 1 fluoro-2-nitro-bcn/\l)-amino|-aeetic acid meth\ I ester
(5.91g, 17.2 mmol) in THF (50 mL) was added lithium hydroxide (4.35 g , 103.6 mmol.) dissolved in
water (17 mL). The reaction was stirred at r.t. overnight. After completion of the reaction, as
confirmed by TLC, the reaction mixture was acidified to pll -I using 10% HCI and crude compound
was extracted with MeOH and CHCL (1:9. 3.\M) mL). Hie combined organic layers were dried o\er
Na2SC>4. The solvents were removed in vacuo to afford 2-{/er/-butoxycarbonyl(4-fluoro-2-
nitrobenzyl)amino)- acetic acid (5.11 g, 90%) as brown thick gel, which was used as such without any
further purification for the next step.
ESIMS (m/z): 328.0 (M-l)
Step 4: Preparation of 2-((2-amino-4-fluorobcnzyl)(/err-butoxycarbonyl)amino) acetic acid
To a solution of [/er/-butoxycarbonyl-(4-niioio-2-nitro-ben/.\l)-amino|-acetic acid (5.11 g. 15 50
mmol) in absolute EtOH (30 mL). was added 10% Pd/C (1.02 g, 20% w/w). The reaction was stirred
at r.t. for 5 h under hydrogen atmosphere. After completion of the reaction, as confiimcd b> I I.C , the
reaction mixture was filtered through eclite bed and was washed with HtOAc (10 mL) and MeOI 1(10
mL). The solvents were removed in vacuo to afford 2-((2-amino-4-fluorobenzyl) (Icrl-
butoxycarbonyl)amino) acetic acid (4.64 g, 99.9%), which was used as such for the next step without
any further purification.
Step 5: Preparation of tert-butyl 8-fiuoro-2-oxo-23-dihydro-17/ benxo[?||l,4] diazcpine-4(57/>-carboxylate
To a solution of [(2-amino-4-fluoro-benzyF) /<'/7-buto\vcarbon>l)-aminoJ-acetic acid (4.64 g. 15.51
mmol) in DCM (60 mL), were added EDC (3.X ? g. 20.1 7 mmol) and HOB'I (2.72 g, 20.17 mmol) at 0
°C. The resulting solution was stirred for 5 nun and DIPLA (8.10 mL, 46.55 mmol) was added. I he
reaction mixture was stirred overnight al r.t under argon. After completion of the reaction as
confirmed by TLC, distilled water (60 mL) was added to the reaction mixture and crude compound
was extracted with DCM (3x50 mL). The combined organic layers were dried over Na7SO(. The
solvents were removed in vacuo to afford the crude compound which was purified by washings with
EtOAc to give pure ter/-butyl 8-fluoro-2-o.\o-2.3-dihydro-17/ benzole][1,4] diazepinc-4(5H)-
carboxylate (2.91 g, 67%) as a white solid.
ESIMS(m/z): 279.5 (M-l)
Step 6: Preparation of 8-fluoro-2-thioxo-l, 2, 3, 5-tctrahydrobenzo|e||l,4]diazepine-4-carboxylic
acid fe/f-butyl ester
To a solution of 8-fluoro-2-oxo-l. 2, 3, 5-tctrahydro-ben/o|c,]ll,4]diazcpinc-4-carboxylic acid tcri-
butyl ester (3.24 g, 11.57 mmol) in toluene (50 mL). was added Lawesson*s reagent (2.33 g. 5 78
mmol) at r.t. The reaction mixture was heated at 90 °C for 30-40 min. After completion of reaction, as
confirmed by TLC, distilled water (50 ml.) was added and the crude compound was extracted with
EtOAc (3x50mL). The combined organic layers were dried over NaiSQj. The solvents were removed
in vacuo to afford the crude compound, which was purified by column chromatography (silica gel. 2:8
EtOAc: Pet. ether) to afford pure 8-fluoro-2-thioxo-l. 2. 3, 5-tetrahydro-bcnzolt>|[l,4|diazepine-4-
carboxylic acid tert butyl ester (2.36 g, 69%) as light \ellovv solid.
ESlMS(nVz): 297.3 (M+l)
Step 7: Preparation of 9-fluoro-477, 677-2, 3, 5, 10A-tctraa/.a-beny.o||az.ulcne-5-carboxylic acid
tert-butyl ester
To a solution of 8-fluoro-2-thioxo-1.2,3,5-tetrahydro-ben/o|c|| l,4|diazepinc-4-carbo.\\lic acid wn-
butyl ester (2.36 g, 7.97 mmol) in dry ben/enc (25 ml), was added formic acid hydrazide dissolved in
5 mL of dry DMSO (1.43 g, 23.91 mmol) at mom temperature. The reaction mixture was heated at 80
°C, using Dean-Stark apparatus for 18 h. After the completion of the reaction, as confirmed bv I LC,
distilled water (40 mL) was added and the crude compound was extracted with diethyl ether (3x40
mL). The combined organic layers were dried over Nla;SO( and concentrated in vacuo to afford the
crude compound which was purified by column chromatography [silica gel, 1.5:8.5 Acetone : Pet.
ether] to afford 9-fluoro-4H, 6H-2, 3, 5. 10/>-tetraa/_a-bcnzo|t'|a/.ulenc-5-carboxylic acid terl-buiyester, (1.455 g, 60%) as a white solid.
ESIMS(m/z): 305.5 (M+l)
Step 8: Preparation of 9-fluoro-5, 6-dihydro-477-2, 3, 5, 10b-tetraaza-benzo[f) azulenc
trifluoroacetate
To a solution of 9-fluoro-4H, 6Z/-2, 3, 5, 10/>-tetraaza-benzo[Vlazulene-5-carboxylic acid tert-buty\ ester, (1.45 g, 4.76 mmol) in dry DCM (15 ml.) was added trifluoroacetic acid (14.30 ml.. 3 mL/mmol) at 0 °C. The reaction mixture was stirred at r.t. for 2 h under argon. After completion of the reaction as confirmed by TLC, MeOH (2xl()ml.) was added and the reaction mixture was evaporated under vacuo to remove excess of solvent and trifluoroacetic acid. The remaining solvents were removed under vacuo and 9-fluoro-5. 6-dih>dro-4H-2. 3, 5, 10b-tetraa/ii-ben/o|c'| a/.ulene trifluoroacetate (1.50 g, 4.71 mmol, 99%) was obtainicd as thick brown jelly. The crude compound was used as such without any further purification for the next step. ESIMS (m/z): 205.2 (M+l), for free amine
Step 9: Preparation of [3-(9-fluoro-4i7,6H-2, 3, 5, 10A tetraaza-benzo[e]-azulen-5-yl)-3-oxo-l-(2, 4, 5-trifluoro-benzyl)-propyl]-carbamic acid-tert-but\\ ester
To a solution of (/?)-3-[(ter/-butoxycarbonyl)amino|-4-(2,4,5-trifluorophenyl)butanoic acid (1.56 g, 4.68 mmol) in dry acetonitrile (15 mL), was added DIP!.A (2.03 inL. 1 1.71 mmol). followed b\ the addition of (benzotriazolyl-l-yloxy)-fm(dimethylaminc) phosphonium hexafluorophosphate (2.27 g, 5.15 mmol) at r.t. 9-fluoro-5, 6-dihydro-4H-2. 3. 5. 10/>-tetraaza-benzo|e]a/.ulene trifluoroacetate (1.56 g, 4.91 mmol), dissolved in dry acetonitrile (lOnil ) and DIPLA (0.98 ml.. 5.63 mmol). was added to the reaction mixture at r.t. The reaction was stirred at 40 "(.' for 48 h. After completion of the reaction, as confirmed by TLC, solvent was removed in vacuo and distilled water (40 mL) was added to the residue. The crude compound was extracted with LtOAc (3x50 mL) and the combined organic layers were dried over Na2SO.j. The solvents were removed in vacuo to afford the crude compound. which was purified by column chromatography (silica gel. 1.2:8.8 MeOH: CHCL) to afford 3-(9-fluoro-4H,6i/-2, 3, 5, 10b tetraaza-benzo[e,|-a/ulen-5-\l)-3-oxo-l-(2, 4, 5-trifluoro-benzyl)-propyl]-carbamic acid-ter/-butyl ester, (2.04 g, 83.9 %), as a white solid solid.
*H NMR (400 MHz, CDC1,) : 8 1.35 and 1 37 (two rotamcric singlets. IH), 2.62-2.93 (m, 2H). 2.95-2.99 (m, 2H), 4.14-4.17 (m, IH), 4.41 (s. 111). 4.57-4.60 (m. 111), 4.76 (s. 111). 4.93 (s. 111). 5.44-5.46 (m, IH), 6.86-6.89 (m, IH), 7.03-7.07 (m. 111). 7 18-7.23 (m. 2H), 7.57-7.60 (m. 111). 8.49 and 8.50 (two rotameric singlets, 111) ESIMS (m/z): 520.5 (M+l)
Step 10: Preparation of trifluoroacetic acid salt of (fl)-3-amino-l-{9-fluoro-4H,6H-2,3,5,10fr-tetraaza-benzo[«]azulen-5-yl)-4-(2,4^-trifluoro-plicnyl)-butan-l-onc
To a solution of [3-(9-fluoro-4H,6H-2, 3. 5. 10b tetraa/a-ben/o[c|-azulen-5-yl)-3-oxo-l-(2. 4. 5-trifluoro-benzyl)-propyl]-carbamic acid-fc#7-but\ I ester. (2.04 g. 3.93 mmol), in dry DCM (12 mL) was added trifluoroacetic acid (11.79 mL, 3 mL/mmol) at 0 °(\ The reaction mixture was stirred at r.t. for 2 h under argon. After completion of the reaction as confirmed by TLC, methanol (2x5mL) was added and the reaction mixture was evaporated under vacuo to remove excess of solvent and trifluoroacetic acid. The remaining solvents were removed under vacuo to obtain trifluoroacetic acid
salt of (/?)-3-amino-l-(9-fluoro^H,6H-2,3.5.10/)-tetraaza-benzo|e]azulen-5-ylH-(2,4,5-trilliioro-
phenyl)-butan-l-one as a white solid (1.82 g, 3.42 mmol, 87%).
'HNMR (400 MHz, CD3OD): 8 2.62-3.15 (m. 4H). 3.87 (bs, 1H), 4.50^.52 (m, 1H), 4.52^.70 (m,
1H), 4.82-4.90 (m, 2H), 7.20-7.34 (m, 3H), 7.58-7.64 (m. 2H), 9.02-9.03 (two rotameric singlets. 1H)
ESIMS (m/z): 420.5 (M+l), Mass of free amine
Example 7: Preparation of trifluroacetic acid salt of (.K)-3-ainino-l-(4iJ,6H-3,5,106-triaza-
benzo[e]azulen-5-yl)-4-(2,4^-trifJuoro-phenyl)-butaii-l-one
(Formula Removed)
Step 1: Preparation of 2-(2-hydroxy-ethylamino)-3^-dihydro-benzo[ff|[l,4Jdiazcpine-4-carboxylic acid terf-butyl ester
2-aminoethanol (3.5 mL, 10 mL/mmol) was added to a solution of ter/-butyl 2-thioxo-2,3-dihydro-lH-benzo[e][l,4]diazepine-4(5H)-carboxylate (100 mg, 0.35 mmol), [prepared as described in Example 6, Step 6] and the reaction mixture was heated to reflux for 24 h under nitrogen atmosphere. The reaction mixture was cooled to room temperature, diluted with ethylacetate (50 ml.) and washed with water (20 mL). The organic layer was separated, dried over anhydrous sodium sulphate and concentrated to give the desired compound as \iscous oil (100.0 mg. 90 %) which was used as such for the next step.
ESIMS (m/z): 328.3 (M+Na), 305.9 ( M-\ 1).
Step 2: Preparation of 4H,6H-3,5,10^triaza-beiizo|e]ttzulene-5-carboxylic acid tert-butyl ester To a solution of DMSO (0.05 mL, 0.82 mmol) in DCM (5.0 ml,) at -78 °C, was added oxalyl chloride (0.04 mL, 0.43 mmol) dropwise. The reaction mixture was stirred at this temperature for 0.5 h. followed by dropwise addition of a solution of 2-(2-hydroxy-ethylamino)-3,5-dihydro-benzo[e][l,4]diazepine-4-carboxylic acid fer/-butyl ester (100 mg, 0.32 mmol) in DCM (1 mL)- The reaction mixture was stirred at -78 °C for 2 h. Ttriethylamine (1.0 mL) was added to the reaction mixture and warmed to room temperature. The crude product was extracted with DCM (20 ml.). The organic layer was washed with water (2*15 mL.). dried over anhydrous Na?SO.| and concentrated in vacuo to afford the crude compound which was purified by column chromatography (silica gel. 8:2 EtOAc:PetEther) to afford 4H,6H-3,5.10/'-triaza-ben/o|t'|a/.ulenc-5-carboxylic acid /e/v-but\l ester (25 mg, 27 %) as a viscous oil. ESIMS (m/z): 301.3 (M+l).
Step 3: Preparation of [3-oxo-3-(4^,6/^-3^,106-triaza-benzo[e]azulen-5-yl)-l-(2,4,5-trifluoro-benzyl)-propyl]-carbamic acid terf-butyl ester.
4H,6H-3,5,10A-Triaza-benzo[elazulene-5-carboxyric acid tert-bu\.y\ ester was dcprotected using
trifluoroacetic acid and the resulting salt was coupled with (/?)-3-[(ferr-butoxycarbonyl)amino|-l-
(2,4,5-trifluorophenyl)butanoic acid using HOB I. LDC and DIPLA to afford [3-oxo-3-(4/A6H-
3,5,106-triaza-benzo[e]azulen-5-yl)-l-(2,4,5-tritluorcvbenzyl)-propyl]-carbamic acid fcrV-butyl ester
as a viscous oil.
ES1MS (m/z): 424.1 (M+Na), 401.4 (M+l).
Step 4: Preparation of trifluroacetic acid salt of 3-amino-l-(4H,6H-3,5,10A-triaza-
benzo[e]azulen-5-yI)-4-(2,4,5-trifluoro-phenyl)-butan-l-one
To a solution of [3-oxo-3-(4/f,6H-3,5,10/)-triaza-benzo[e]azulen-5-yl)-l-(2,4,5-trifluoro-benzyl)-
propyl]-carbamic acid tert-buty\ ester (30 mg, 0.06 mniol) in DCM (3 mL), was added trifluoroacetic
acid (0.18 mL, 3 mL/mmol)) . The reaction mixture was stirred at r.t. for 1 h. After the completion of
the reaction as confirmed by TLC, excess of trichloroacetic acid and DCM were removed in vacuo lo
afford a gummy solid which was crystallised from he\ane to afford trifluroacetic acid salt of 3-aniino-
l-<4H,6H-3,5,10*-triaza-benzo[e]azulen-5-yl)-4-{2.4.5-trinuoro-phcnyl)-butan-l-one (30 mg, 98%).
'H NMR (400 MHz, MeOD): 8 2.72-2.99 (in. 2H), 3.00-3.17 (m, 2H), 3.80-3.98 (m. 1H), 4.51-4.70
(m, 2H), 4.72-5.08 (m, 2H), 7.12-7.27 (m, 1H). 7.28-7.42 (m, 1H), 7.45-7.81 (m, 511), 7.82-8.12 (in.
1H)
ESIMS (m/z): 424.1 (M+Na), 401.4 (M+l)
Example 8: Preparation of trifluroacetic acid salt of (/?)-3-amino-l-(4H,6H-l,2£,10£-tetraaza-
benzo[«]aznlen-5-yI)-4-<2,4^-trifluoro-phenyl)-butan-l-one
(Formula Removed)
Preparation of (2-Azido-benzyl)-carbamic acid terf-butyl ester
To a solution of (2-amino-benzyl)-carbamic acid /tW-butyl ester Iprcpared from 2-aminobenzylamine and di-terf-butyl dicarbonate], (100 mg, 0.45 mmol) in acetic acid (2.46 mL), was added water (26 mL). Reaction mixture was cooled to 0 °C and a solution of sodium nitrite (62 mg, 0.9 mmol) in water (4.3 ml) was added. The reaction mixture was stirred for 30 min at the same temperature followed by addition of sodium azide (64 mg, 0.99 mmol). The reaction mixture was stirred at 0 °C for 30 min. When the reaction was complete as confirmed by TLC. reaction mixture was quenched by dropwise addition of 10 N NaOH (5.0 mL). The crude compound was extracted with ethylacetate (2 *10 mL). Organic layer was separated, washed with brine, dried over Na^SO-t and the solvents were removed in vacuo to afford (2-azido-benzyl)-carbamic acid ti'rt-b\.\\\\ ester (105 mg, 96 %) which was used as such without any further purification for the next step. ESIMS (m/z): 271.3 (M+Na), 249.1 (NM 1).
Step 2: Preparation of (2-azido-beiizyl)-prop-2-ynyl-carbamic acid tert-butyl ester
Sodium hydride (5.0 mg, 0.120 mmol, 60% suspension in mineral oil) was washed with hexane (2 mL) in a flame dried round bottomed flask under nitrogen atmosphere. To the resulting free floating powder, was added a solution of (2-azido-ben7yl)-carbamic acid tcrt-butyl ester (20 mg, 0.08 mmol) in dry DMF (1 mL) at 0 °C. The reaction mixture was Mined at this temperature tor 30 min. Propargx I bromide (0.021 mL, 0.24 mmol) was added and the reaction mixture was stirred at r.t. for 2 h. Alter the completion of reaction as confirmed by I'LC . crude product was extracted with ethylacetatc (10 mL). The organic layer was washed with water (2*5 ml-). Organic layer was dried over Na.SO, and concentrated in vacuo to afford the crude compound which was purified by column chromatography (silica gel; 1:9 EtOAc:Pet.Ether) to afford (2-azido-benzyl)-prop-2-ynyl-carbamic acid ff/7-butyl ester (20 mg, 87%) as a viscous oil. ESIMS (m/z): 309.2 (M+Na), 287.4 (M+ 1).
Step 3: Preparation of 4/J,6H-l,2,5,10A-tetraaza-benzo|«|a;eulene-5-carboxylic acid /erf-butyl ester
Toluene (3 mL) was added to (2-azido-ben/yl)-prop-2-ynyl-carbamic acid terl-buty\ ester (100 mg, 0.34 mmol) and the resulting mixture was heated at 100 °C overnight. When the reaction was complete as confirmed by TLC\ solvent was removed in vacuo to afford 4H.6H-1.2.5.1()/>-tcuaa/a-benzo[e]azulene-5-carboxylic acid tert-buly\ ester (92 mg. 92%) which was used as such without an\ further purification for the next step. LSIMS (mV): 309.4 (M i Na). 287.1 (Mi I). Step 4: Preparation of |3-oxo-3-(4H,6H-lr2,5,10A-tetraaza-benzo|f|azulen-5-yl)-l-(2,4^-trifluoro-benzyl)-propyl]-carbainic acid tert-butyl ester
4H,6^/-l,2,5,106-Tetraaza-benzo[e]azulene-5-earbox\lie acid /cr/-butyl ester was deprotected using trifluoroacetic acid and and the resulting salt was coupled with (/0-3-|(ter/-butoxycarbonyl)amino|-4-(2,4,5-trifluorophenyl)butanoic acid using HOBt. EDC and DIPLA. The desired compound was obtained as viscous oil. ESIMS (m/z): 524.9(M+Na), 502.5 (MM).
Step 5: Preparation of trifluroacetic acid salt of (/?)-3-amino-l-(4H,6H-3,5,10A-triaza-benzo[e]azulen-5-yl)-4-(2,4,5-trifliioro-phenyl)-biitan-1 -one
To a solution of [3-oxo-3-(4H,6H-l,2,5,10A-tctraaza-ben/o|c|a/ulen-5-yl)-l-(2,4.5-lrilluoro-ben/> I)-propyl]-carbamic acid /er/-butyl ester (30 mg. 0.06 mmol) in DC'M (3 mL), was added trifluoroacetic acid (0.18 mL, 3 mL/mmol). The reaction mixture was stirred at r.t. for 1 h. When the reaction was complete as confirmed by TLC, excess of trilluoroacetie acid and IX'M were removed in vacuo to afford a gummy solid which was crystallised from hexane to afford trifluroacetic acid salt of (^)-3-amino-l-(4^/,6H-3,5,10b-triaza-benzo[^Jazulen-5-\l)-4-(2.4.5-trilluoro-phenyl)-butan-l-one (30 mg. 98%)
!H NMR (400 MHz, MeOD): S 2.52-2.83 (m, 2H), 2.85-2.88 (m, 2H), 3.59-3.65 (m, 1H), 4.51 (s,
1H), 4.58 (s, 1H), 4.72-4.78 (m, 2H), 7.12-7.17 (m. 1H), 7.25-7.29 (m, 1H), 7.53-7.60 (m, 2H), 7.62-
7.67 (m, 1H), 7.90 (s, 1H), 7.91-7.99 (m, 1H):
ESIMS (m/z): 424.4 (M+Na), 402.4 (NH 1)
Example 9: Preparation of trifluroacetic acid salt of (^)-3-amino-l-[3-(4-fluoro-phenyl)-4H,6H-
1^2^,10b-tetraaza-benzo[f]azulen-5-yl]-4-(2,4^-trifluoro-phenyl)-bntan-l-onc
(Formula Removed)
Step 1: Preparation of 3-(4-fluoro-phenyl)-prop-2-yn-l-ol
To a mixture of 4-fluoroiodobenzene (710.0 mg. 3.82 mmol), bistriphenylphosphine palladium chloride (27.0 mg, 0.038 mmol), Cul (3.64 mg. 0.019 mmol) and propargyl alcohol (214.0 mg, 3.82 mmol), was added diisopropylamine (6.1 ml.. 1.6 niL/mmol) under nitrogen atmosphere. The reaction mixture was stirred at r.t. for 6 h. After the completion of the reaction as confirmed by TLC. the crude product was extracted with ethylacetate (50 ml.) The organic layer was washed with 10% IK I ( 20 mL), dried overNajSO^ and concentrated in vacuo to afford the crude compound which was punllcd by column chromatography (silica gel, 2:8 Lt()Ac:Pet.l-.ther) to afford 3-(4-fluoro-phenyl)-prop-2-yn-l-ol (366.0 mg, 64%) as a viscous oil.
Step 2: Preparation of l-(3-bromo-prop-l-ynyl)-4-fluoro-benzene
To a solution of 3-(4-fluoro-phenyl)-prop-2-\ii-l-ol (360 mg. 2.4 mmol) in dry DCM (5 mL) at 0 °C. was added triethylamine (0.7 mL, 3.6 mmol) under nitrogen atmosphere. The reaction mixture was stirred for 15 min. followed by dropwise addition of methanesulfonyl chloride (0.3 mL, 3.6 mmol). The mixture was stirred at the same temperature for 30 min. After the completion of the reaction as confirmed by TLC, the crude product was extracted with DCM (10 ml.). The organic la\er was washed with water (10 mL), separated, dried o\er Na^SO) and concentrated in vacuo to afford the mesylated product as viscous oil which was used as such for next step, lo the resulting compound (600 mg, 21.04 mmol) in dry THF (20 ml.) at 0 °C. was added solid Lithium bromide (1.8 g. 21.63 mmol) under nitrogen atmosphere. The reaction mixture was stirred at r.t. for 2 h. Attei the completion of the reaction, as confirmed b\ I I.C. water (20 ml.) was added to the reaction mixture and the crude compound was extracted with ethylacetate (20 ml.). The organic layer was separated, dried over Na2SC>4 and concentrated in vacuo to afford the crude compound, which was purilled by
column chromatography (silica gel, 2:8 LtOAcil'et.l-.ther) to afford l-(3-bromo-prop-l-yn\i)-4-fluoro-benzene (460 mg, 82.5 %) as a viscous oil. ES1MS (m/z): 214.5 (M+2)
Step 3: Preparation of 3-(4-fluoro-phenyl)-4H,6H-l^^,106-tetraaza-bciizo[e]azuleDe-5-carboxylic acid ferf-butyl ester
Sodium hydride (80.0 mg, 2.0 mmol, 60% suspension in mineral oil) was washed with hexane (2 mL) in a flame dried round bottomed flask under nitrogen atmosphere. To the resulting free floating powder, was added a solution of (2-azido-benzyl)-carbamic acid /er/-butyl ester (250 mg, 1.0 mmol) in dry DMF (3 mL) dropwise at 0 °C under nitrogen atmosphere. The reaction mixture was stirred at this temperature for 30 min followed by dropwise addition of a solution of l-(3-bromo-prop-l-ynyl)-4-fluoro-benzene (318.0 mg, 1.50 mmol) in dry DMI (1.5 mL). The reaction mixture was stirred at r.t. for 2 h. After the completion of the reaction as confimied by TI.C, the crude compound was extracted with ethylacetate (2x10 mL). 1 he combined organic layer was washed with water (2-10 mL). Organic layer was separated, dried over Na>S()| and concentrated in vacuo to afford the crude compound which was purified by column chromatography (silica gel. 1:9 LtO Ac: Pet. Ether) to afford 3-(4-fluoro-phenyl)-4H,6H-l,2,5,10b-tetraa/ii-ben/o|e|a/.ulene-5-carboxylic acid terf-butyl ester (70 mg, 12.2 %) as a viscous oil. ESIMS (m/z): 381.6 (M+l).
Step 4: Preparation of [3-[3-<4-fiuoro-phenylHff,6H-l^^,106-tetraaza-benzo[e]aznlen-5-yl]-3-oio-l-(2,4^-trifluoro-benzyl)-propyl]-carbaniic acid tert-buiyl ester
3-(4-Fluoro-phenyl)-4H,6H-l,2,5,10ft-tetraa/a-bcnzo|e|azulcne-5-carboxylic acid /e/7-butyl ester was deprotected using trifluoroacetic acid and the resulting salt was coupled with (R)-3-\(lert-butoxycarbonyl)amino|-4-(2,4,5-trifluorophen\ l)butanoic acid using HOB I, l.DC and DIP1 A. Hie desired compound was obtained as a viscous oil. ESIMS (m/z): 619.1 (M+Na), 596.9 (M+l).
Step 5: Preparation of trifluroacetic acid salt of (^)-3-amino-l-[3-{4-fluoro-phenyl)-4y/,6H-l^^,106-tetraaza-benzo[«]azulen-5-yi]-4-(2,44»-trifluon)-phcnyI)-butan-l-one To a solution of [3-[3-(4-fluoro-phenyl)-4H.6/y-l.2.5.1()A-tctraa/a-benzo|c|a/ulen-5-yl|-3-oxt)-l-(2,4,5-trifluoro-benzyl)-propyl]-carbamic acid tert-buly\ ester (28.0 mg, 0.047 mmol) in 1XM (2 mL), was added trifluoroacetic acid (0.18 mL, 3mL/mmol). The reaction mixture was stirred at r.t. for 1 h. After the completion of the reaction as confirmed by TLC, excess of trifluoroacetic acid and DCM were evaporated in vacuo to afford a gumm> solid which was crystallized from hexane to afford trifluroacetic acid salt of (/?)-3-amino- l-|3-(4-fluoro-phenyl)-4H,6H-1,2,5,1 Ofc-tetraaza-benzo[e]azulen-5-yl]-4-(2,4,5-trifluoro-phen\ h-butan- I -one (25 mg. 87 %).
'H NMR(400 MHz, MeOD): 6 2.67-2.81 (m. 4H). 3 17^.53 (m. 111). '1.56-1.61 (m. 111). 7 1 I i \() (m, 4H), 7.58-7.79 (m, 5H), 7.98-8.02 (m, 1II) ESIMS (m/z): 496.7 (M+l)
Example 10: Preparation of triflnroacetic acid salt of 5-|(^)-3-amino-4-(2,4,5-trifliioro-phenyI)-butyryl]-5,6-dihydro-4H-2^,106-triaza-benzo|e|azulenc-3-carboxylic acid ethyl ester
(Formula Removed)
Step 1: Preparation of 4H,6H-2^,106-triaza-benzo|f|a/.ulene-3,5-dicarboxylic acid 5-tert-butyl ester 3-ethyl ester
To a solution of 2-oxo-l,2,3,5-tetrahydro-benzo[e|[ l,4]diazepine-4-carboxylic acid ter/-butyl ester (10.0 g, 38.16 mmol) in dry THF ( 300 ml.) at 0 °C, was added potassium terf-butoxide (6.4 g, 56.76 mmol) under nitrogen atmosphere and the reaction mixture was stirred at this temperature for 30 min followed by dropwise addition of diethylchlorophosphate (1 1 ml,. 76.33 mmol). The reaction mixture was stirred at 0 °C for 40 min. This solution was transferred via cannula to a suspension of potassium te/7-butoxide (12.86 g, 114.56 mmol) and eth\ I isocvaiioacctatc (1 1.06 ml., 102.65 mmol) in dr\ 11II (150 mL), kept in a separate flask at 0 C under nitrogen atmosphere. When the addition was complete, the reaction mixture was allowed to come to r.t. and stirred for 45 min. during which the reaction was complete as confirmed by TI.C Reaction mixture was cooled to 0 °C and quenched with 10 % acetic acid solution and stirred for 20 min. The crude compound was extracted with ethylacetate. The organic layer was washed with water, separated, dried over Na.^SOi and concentrated in vacuo to afford the crude compound which was purified by column chromatography (neutral alumina using neutral alumina, 3:7 LtOAciPet.Kther) to afford a viscous gel which was further purified by crystallisation from diethyl ether and hexanc to afford 4H,6H-2,5.10/>-triaza-benzo[e]azulene-3,5-dicarboxylic acid 5-/t'/7-but\ 1 ester 3-ethyl ester a white solid (4.4 g, 33%)
ESIMS (m/z): 380.6 (M+Na), 358.3 (M+l).
Step 2: Preparation of 5,6-dihydro-4H-2,5,10A-tria/.a-ben/.o||a/.ulcnc-3-carhoxylic acid ethyl ester trifluoroacetate
To a solution of 4H,6H-2,5,10ft-Triaza-ben/.o|c|a/.ulcnc-3.5-dicarboxylic acid 5-/cr/-butyl ester }-ethyl ester (300 mg, 1.14 mmol) in DCM (30 ml.), was added trifluoroacetic acid ( 3.42 ml.. 3 ml/mmol) and the reaction mixture was stirred for 2 h. After the completion of the reaction as confirmed by TLC, excess of trifluoroacetic acid and IX'M were removed in vacuo to afford x6-dihydro-4.r7-2,5,106-triaza-benzo[e]azulene-3-carboxylic acid ethyl ester trifluoroacetate (295 mg, 93 %) as a gummy solid which was used without am further purification for the next step. ESIMS (m/z): 280.7 (M+Na), 258.3 (Ml 1).
Step 3: Preparation of (/f)-5-[3-tert-butoxycarbonylamino-4-(2,4,5-trifluoro-phcnyl)-butyryl|-5,6-dihydro-4/f-2^,106-triaza-benzo[e]azulene-3-carboxylic acid ethyl ester
To a solution of (R)-3-[(ter^butoxycarbon\l)amino|-4-(2.4,5-triiluorophenyl)butaiK>ic acid (80 mg. 0.26 mmol) in dry DCM ( 15 mL), was added IIOB1 (42 mg. 0.31 mmol), EDC (60 mg, 0.31 inmol) and DIPEA (0.21 mL, 1.21 mmol) and the reaction mixture was stirred for 10 min. 5,6-Dihydro-4H-2,5,10A-triaza-benzo[e]azulene-3-carboxylic acid ethyl ester trifluoroacetate ( 98 mg, 0.26 mmol) in DCM (5 mL) was added and the reaction was stirred at r.t. overnight under nitrogen atmosphere. After the completion of the reaction, as confirmed b\ TLC. the crude product was extracted with DCM (10 mL) and washed with 10% HCI solution (10 ml.) and saturated sodium bicarbonate solution (10 mL) sequentially. The organic layer was separated, washed with water, dried over NaSO. ;ind concentrated in vacuo to afford the crude compound, which was purified by column chromatography (silica gel, 3:7 EtOAc:Pet.Ether) to afford (/£)-5-|3-/t^buU)xycarbonylamino-4-(2.4.5-trilliioro-phenyl)-butyryl]-5,6-dihydro-4H-2,5,10/>-tria/a-benzo|t'|a/.ulene-3-carboxylic acid ethyl ester (105 mg, 77 %) as a viscous oil. ESIMS (m/z): 596.0 (M+Na), 573.9 (M-< I)
Step 4: Preparation of trifluroacetic acid salt of 5-|(/c>3-amino-4-(2,4,5-trifluoro-phenyl)-butyryi]-5,6-dihydro-4H-2^,10A-triaza-benzo|e|azulene-3-carboxylic acid ethyl ester
To a solution of (^?)-5-[3-/crt-butoxycarbonylamino-4-(2.4,5-tritluoro-phenyl)-butyryl]-5,6-dihydro-4H-2,5,10&-triaza-benzo[e]azulene-3-carboxy lie acid ethyl ester (100 mg, 0.174 mmol) in IX'M (5 mL), was added trifluoroacetic acid (0.52 ml . ? inL'mmol). The reaction mixture was stirred at r.t. for 1 h. After completion of the reaction, as confirmed by 'ILC, excess of trifluoroacetic acid and DCM were evaporated in vacuo to afford a gummy solid which was crystallised from hexane to alford trifluroacetic acid salt of 5-[(/?)-3-amino-4-(2.4.5-trit1uoro-phenyl)-butyryl]-5.6-dihydro-4H-2.5.l0/)-triaza-benzo[e]azulene-3-carboxylic acid ethyl ester (80 mg. 79%).
'HNMR(400 MHz, MeOD): 6 1.39 (t,./ 4.0 HA 311), 2.80-3.06 (m, 2H), 3.05-3.14 (m. 211). 5.90 (m, 1H), 4,35 (q,y= 4.0 Hz, 2H), 4.43-4.62 (m, 3H), 4.77-5.01 (m. 1H), 7.19-7.26 (m, 1H), 7.3 1-7.37 (m, 1H), 7.51-7.61 (m, 2H), 7:62-7.72 (m. 211). 8.25 (s. 1H) ESIMS (m/z): 495.8 (M+Na), 473.5 (M t 1)
Example 11: Preparation of trifluroacetic acid salt of 5-[(/?>-3-amino-4-{2,4,5-trifluoro-phenyl)-buryryl]-5,6-dihydro-4^-2,5,10A-triaza-bi;n/.(»|^|a/.ulene-3-<'arb<)xylic acid
(Formula Removed)
Step 1: Preparation of 5-[3-ter/-butoxycarbonylamino-4-(2,44>-trifluoro-phenyl)-butyr>l|-5,6-dihydro-4/7-2,5,101>-triaza-benzo[£]aziilene-3-carboxylic acid:
To a solution of 5-L3-/er/-butoxycarbonylamino-4-<2.-l;5-tri!liioro-phcnyl)-butyr>'l|-5.6-dihytlio-MI-2,5,10ft-triaza-benzo[e]azulene-3-carboxylic acid eth\ I ester ( 100 mg. 0.17 mmol) in IHl- (5 nil.) and water (5 mL), was added lithium hydroxide (43 mg, 1.03 mmol) . The reaction mixture was stirred at r.t. overnight. After the completion of the reaction as confirmed by TLC. the reaction mixture was cooled to 0 °C and neutralized with 10 % HC1 solution to pH~4. The compound was extracted with ethylacetate (15 mL). The organic layer was washed with water, separated, dried over Na2S04 and concentrated in vacuo to afford 5-|3-/er^-butoxycarbonylamino-4-(2,4,5-tritluoro-phenyl)-butvryl]-5,6-dihydro-4H-2,5,10ft-tria/ji-ben/o|t'|azulene-3-carboxylic acid (85 mg, 89 %) as a viscous gel which was used without further purification for the next step. ESIMS (m/z): 568.0(M+Na), 546.0(M42)
Step 2: Preparation of trifluroacetic acid salt of 5-|(/?)-3-amino-4-(2,44»-trifluoro-phenyl)-butyryl]-5,6-dihydro-4/f-24>,106-triaza-b*n/.»>|<'|azulcnc-3-carbo\ylic acid
To a solution of (/?)-5-[3-te/7-butoxycarbon\ lamino-4-( 2.4.5-trilluoro-phenyl)-butyryll-5.6-dih\ dro-4H-2,5,10£-triaza-benzo[e]azulene-3-carbox>lic acid (35 mg. 0.06 mmol) in dry DCM (5 mL), was added trifluoroacetic acid (0.18 mL, 3 mL/mmol). flic reaction mixture was stirred at r.t. for I h. After the completion of the reaction as confirmed b> 1 LC. excess of trifluoroacetic acid and DCM were evaporated in vacuo to afford a gumm\ solid which was crystallised from hexanc to afford trifluroacetic acid salt of 5-[(/?)-3-amino-4-(2,4.5-trifluoro-phenyl)-butyryl]-5,6-dihydro-4f/-2,5,10^-triaza-benzo[e]azulene-3-carboxylic acid (28 mg. 78%).
'H NMR (400 MHz, DMSO): S 2.80-2.99 (m. 4H). 3.72-3.80 (m, 2H), 4.31-5.01 (m, 3H), 7.49-7.54 (m, 3H), 7.71-7.73 (m, 2H), 7.92-7.95 (m, 211) ESIMS (m/z): 443.8 (M-l)
Example 12: Preparation of trifluroacetic acid salt of (/?)-3-amino-l-|3-(p>rroliiliiu--l-carbonyl)-4if,6fl-2^,10A-triaza-benzo[c]azulcn-5-yl|-4-(2,4^-trifluoro-phcnyl)-butan-l-onc
(Formula Removed)
Step 1: Preparation of (^)-|3-oxo-3-|3-(pyrrolidinc-l-carbonyl)-4H,6H-2^,106-triaza-benzo|«]azulen-5-yl]-l-(2,4^-trifluoro-benzyl)-propyl|-carbamic acid tert-butyi ester
To a solution of (/?)-5-[3-rer/-butoxycarbonylamino-4-(2.4.5-trifluoRvphenyl)-butyryl]-5,6-dihydro-4H-2,5,10/»-triaza-benzo[e]a/.ulenc-3-carbo\> lie acid ((>() mg. 0 1 1 mmol) in dr\ IX'M (15 ml ). was added HOBT (19 mg, 0.14 mmol), EDC (27 nig. 0.1 I mmol) and Dll'l-A (0.08 ml . 0.16 niinoli and the reaction mixture was stirred for 10 min. l'\rrolidinc (0.02 mL, 0.24 mmol) was added and the
reaction was stirred at r.t. overnight under nitrogen atmosphere. After the completion of the reaction, as confirmed by TLC, the crude product was extracted with lX'M (15 mL) and washed with 10% HC1 solution (10 mL) and saturated sodium bicarbonate solution (10 mL). The organic layer was separated washed with water, dried over Na>SOj and concentrated in vacuo to afford the crude compound. which was purified by column chromatography (silica gel, 3:7 to 9:1 l',t()Ac:Pel.Lllier) to afford ( R)-[3-oxo-3-[3-(pyrrolidine-l-carbonyl)-4M6H-2,5,10/>-triaza-bcnzo|e]azulen-5-yl]-1 -<2.4,5-trifluoro-benzyl)-propyl]-carbamic acid /er/-butyl ester CO mg. 83 %) as a solid. ESIMS (m/z): 621.0 (M+Na), 599.0 (MH)
Step 2: Preparation of trifluroacetic acid salt of (/{)-3-amino-l-[3-(pyrrolidine-l-carbonyl)-4i/,6i/-2^,10fr-triaza-benzo[e]azulen-5-yl)-4-(2,4^-trifluoro-phenyl)-butan-l-one To a solution of (^?)-[3-oxo-3-[3-(pyrrolidine-l-carbonyl)-4^/,6H-2,5,106-triaza-benzo[eJazulen-5-yl|-H2,4,5-trifluoro-benzyl)-propyl]-carbamic acid /c/7-butyl ester (45 mg, 0.08 mmol) in DCM (5 mL) was added trifluoroacetic acid (0.24 mL. 3 ml. mmol). 1 he reaction mixture was stirred at r.t. for 1 h. After the completion of the reaction, as continued b> I l.C. excess of trilluoroacetic acid and DCM were evaporated in vacuo to afford a gumnn solid which was crystallized from hexane to afford trifluroacetic acid salt of (/i)-3-amino- l-|3-(p\rrolldinc-l-earrH)nyl)-4H.()H-2.5.10/'-t^a/a-benzo[e]azulen-5-yl]-4-(2,4,5-trifluoro-phenvl)-blltan-l-onc (40 mg, 86%)
'H NMR(400 MHz, MeODM 1-98-2.02 (m. 411). 2.97-3.1 1 (m. 4H), 3.62 (t../ 6.0 Hz. 2H). V92-4.05 (m, 3H), 4.41-4.87 (m, 411), 7.19-7.26 (m. 111). 7.31-7.37 (m. 111). 7.51-7.61 (m. 211). 7.62-7.72 (m,2H),8.17(s, 1H) ESIMS (m/z): 520.1 (M+Na), 498.6 (M+1)
The compounds listed in Tables 6 to 9 were prepared essentially following the procedures described for Examples 1 to 12:
Table 6
(Table Removed)
In vitro Studies Protocol for in-vitro DPP-IV assay
DPP-IV activity was determined by the rale o\ cleavage of />-nitro aniline (/;NA) from synthetic substrate GIycine-Proline-/?NA. The assa\ was conducted b\ adding 1 ug of porcine kidney or human recombinant DPP-IV enzyme (Sigma Aldrich. USA) in 100 uL of the assay buffer (100 mM Tris. pH 7.4, 140 mM NaCI, 10 mM KC1, 1% BSA) to 96 well C'ellstar Hat bottom microtitrc plates (Greiner Bio-one, Germany). The reaction was initiated by adding 80 u,L of 500 uM substrate Gly-Pro-pNA. The incubation was carried out in the kinetic mode at 30 "C for 30 min. Absorbance was measured using Synergy HT Biotek Multiplate reader at 410 nm. lest compounds and solvent controls were added as 10 u.L additions. A standard curve of free />-nitro aniline (pNA) was generated using 0-2000 |iMpNA in the assay buffer. In addition DPP-IV acti\it\ was also determined by using Flourogenic substrate (Gly-Pro-AMC) using human recombinant DPP-IV en/sine (Sigma Aldrich l!SA). Tests for ICJQ studies: Test compounds dissol\ed in DM SO at 9-10 dillercnt concentrations were tested in triplicates along with the solvent control and blank samples. %age inhibition was calculated at each concentration with respect to the soKent control (no test compound added). IC\(, values were calculated from 3 experiments using the Graph Pad Prism or Sigma Stat software.
Table 10
(Table Removed)
X: 0-49%) DPP-IV inhibition at lOOnM Y: 50-100% DPP-IV inhibition at lOOnM
Table 11
(Table Removed)
In vivo Studies Oral glucose tolerance test (OGTT) in C57B176J mice
The oral glucose tolerance test (OG I 1 ) measures the bod\'s abilit\ to use ylueose that is ihe body's main source of energy. OG 1 "I" serves as a priman m-vivo screen to select efllcaeious test compounds for their antidiabetic activity. Compounds were formulated in 0.25% CMC with a drop of tween 80 (optional). C57BL/6J male mice (8-9 weeks) were fasted overnight and randomi/ed into different groups (n=6) on body weigh basis. At lis min blood was collected from each group for glucose estimation. At To compounds or vehicle were administered with simultaneous administration of glucose (2g/kg p.o.) to each group. RO water was administered to no glucose control group. Mood samples were collected from retro-orbital plexus at 15. 30. 60 and 120 min post dosing for glucose estimation. The AUC for glucose was calculated to get the reduction in glucose excursion.
Table 12
(Table Removed)
We claim:
A compound of Formula I,
(Formula Removed)
or its pharmaceutically acceptable derivatixes. tautomeric forms, stereoisomers including R and S isomers, polymorphs, prodrugs, metabolites, salts or solvates thereof, wherein:
Ar represents aryl which may be phenyl: which may further be unsubstituted or may be optionally substituted at any available position by one or more substituents selected from but not limited to halogen, CN, hydroxyl, NH2, C1-12 alkyl or C1-12 - alkow: R1 is selected from the group consisting of but not limited to (CH2)nCONRaRb, (CH2)„COOR', (CH2)nNRaRb, (CH2)nNRaCORb, (CH2)nC( I )R" (wherein L is O or S), (CH2)1ORa (wherein each methylene group may be substituted by one or more halogen atoms). -(C'O)Ra. -(C())NK''R". hydrogen, C1-12 alkyl, C2-12 alkenyl. C1-12 alkynyl C1-12 haloalkyl. C2-12 haloalkein 1. C L. haloalkynyl, Cvs cycloalkyl, heterocyclyl. aryl. heteroaryl. (CH2)n-cycloalkyl. (CH2)n-heteroc\cl\l. (CH2)n-aryl, (CH2)„-heteroaryl, each of which may be optionally substituted at any available position by one or more substituents selected from but not limited to hydrogen, halogen. CN. C|-i2 alkyl, C2-12 alkenyl, C2-12 alkynyl. C1-12 alkow. C1-12 haloalkvl. C1-12 haloalkoxy. C2-12 haloalkein I. C2-1 haloalkynyl, C1-12 alkylcarbonyl. C1-12 alkoxycarbonyl, oxo, -OR', -SRa. -NO2 -NR'R', N(Ra)(CO)Rb, N(Ra)(CO)ORb, N(Ra)(CO)NRaRb. -(CO)Ra. -(CO)NRaRb, -O(CO)Ra, -O(CO)NRaRb, -COORa, C3-8 cycloalkyl, S(O)mRa, SO2NRaRb : cycloalkyl which may be optionally substituted at any available position by one or more substituents independently selected from R' or Rc ; aryl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc; heteroaryl which may be optionally substituted at any available position by one or more substituents independent!}, selected from R' or RL : or heterocyclyl which ma\ be optionally substituted at any available position by one or more substituents independent!} selected from Rc or Rc; R2 and R3 together represents a single oxygen or sulpluii' atom which is linked to the dia/.epine ring b\ a double bond; or R1 and R2 together forms a double bond in the diazepine ring and R' represents the group -NRaRb; or R1 and R3 together with the nitrogen atom to which R1 is attached forms a heterocyclic or heteroaryl ring which ma} additionally contain from one to three heteroatoms independently selected from O, S and N: the ring formed may optionally be substituted with one or more substituents selected from Rc or Rc and R represent hydrogen or a double bond;
R4 and R5 are independently selected from the group consisting of hydrogen, halogen, CN, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C1-12 alkoxy, C1-12 haloalkyl. C1-12 haloalkoxy, C2-12 haloalkenyl. C1-12 haloalkynyl, Cri2 alkylcarbonyl, C1-12 alkoxycarbonyl, -OR". -SRa, -NO2,-NRaRb, N(Ra)((O)Rh. N(Ra)(CO)ORb, N(Ra)(CO)NRaRb, -(CO)R9. ((O)NR'Rb, -O(CO)Ra, -O(CO)NR'Rb, -COOR", C3-8 cycloalkyl, S(O)mRa, SO2NRaRb ; cycloalkyl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc; aryl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc; heteroaryl which may be optionalK substituted at any available position by one or more substituents independently selected from Rc or Rc ; or heterocyclyl which may be optionally substituted at any available position by one or more substituents independently selected from R' or
Rc;
R6 and R7 are independently selected from the group consisting of hydrogen, halogen, CN, C1-12 alkyl, C2-12 alkenyl, C2.]2 alkynyl, C1-12 alkoxy. C1-12 haloalkyl. C1-12 haloalkoxy, C2-12 haloalkenyl. C1-12 haloalkynyl, C1-12 alkylcarbonyl, C,-12 alkoxycarbonyl. -OR', -SRa, -NO,. -NRaRb, N(Ra)(CO)R", N(Ra)(CO)ORb, N(Ra)(CO)NRaRb, -(CO)Ra. -(CO)NRaRb, -O(CO)Ra -O(CO)NRaRb, -COORa, C3-8 cycloalkyl, S(O)mRa, SO2NRaRb ; cycloalkyl which may be optionally substituted at any available position by one or more substituents independently selected from RL or RL; aryl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc; heteroaryl which may be optionally substituted at any available position by one or more substituents independently selected from RL or RL ; or heterocyclyl which may be optionalK substituted at any available position by one or more substituents independently selected from W or Rc;
R8 is independently selected from hydrogen, halogen. CN, C1-12 alkyl, C1-12 haloalkyl, C1-12 alkoxy, C1-12 haloalkoxy, C2-12 haloalkenyl, C1-12 alkylcarbonyl. C1-12 alkoxycarbonyl, -ORa. -SRa, -CF3 -OCF3, -NO2, -NRaRb, N(Ra)(CO)Rb, N(R')(CO)ORb. N(Ra)(CO)NRaRb, -(CO)Ra. -(CO)NRaRb. -O(CO)Ra, -O(CO)NRaRb, -COORa, C3-6 cycloalkyl, S(O)mRa, SO2NRaRb; cycloalkyl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc; aryl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc ; heteroaryl which may be optionally substituted at any available position by one or more substituents independently selected from Rc or Rc : or heterocyclyl which may be optionally substituted at any available position by one or more substituents independently selected from Ra or Rb :
Ra and Rb are independently selected from Imlrogen. C1-12 alkyl, C1-12 alkenyl. C2-12 alknyl C1-12 haloalkyl, C2-12 haloalkenyl, C2-12 haloalkynyl, C3-8 cycloalkyl, heterocyclyl, aryl, heteroaryl, (CH2)n-cycloalkyl, (CH2)n-heterocyclyl. (CH2)n-aryl. (CH2)n-heteroaryl; each of which may be optionally substituted with halogen, hydroxyl C1-10 alkyl, C2-12 alkenyl, C2-12 alkynyl, C1-10 alkoxy. C1-12 alkylcarbonyl, C1-12 alkoxycarbonyl. C3-8 cycloalkyl, C1-12 haloalkyl, C1-12 haloalkoxy. C2-12
haloalkenyl, aryl, heterocyclyl, heteroaryl. (CH:)„-aryl. (ClF)„-heterocyclyl. (CH2)„-heteroaryl, (CH2),,-cycloalkyl, oxo, -CN, OR", NO.. NK"R'". N(R9)(CO)R10. N(R9)(CO)OR". N(R9)(CO)NR9R10, -C(=L)R9 (wherein I is () or S). -(CO)NR9R10. -O(CO)R9. -O(CO)NR9R10. COOR9, -SR9, S(O)mR9, SO2NR9R10; SO3H. NHSO.R9. P(O)R10; or Ra and Rb may be joined together along with the nitrogen atom to which they arc attached to form a heterocyclic or heteroaryl ring which may additionally contain from one to three heteroatoins independent!} selected from O, S and N, the ring formed may optionally be substituted with one or more substituents selected from hydrogen, halogen. C1-12 alkyl. C2-12 alkenyl, C2-12 alkynyl. C1-12 haloalkyl, C2-12 haloalkenyl. C2-12 haloalkynvl, C3-8 cycloalkyl, heterocyclyl, aryl, heteroaryl, (CH2)n-cycloalkyl, (CH2)n-heterocyclyl, (CH )n-aryl 1. (ClF)„-heteroaryl, C\-\?. alkylcarbonyl. Cn2 alkoxycarbonyl, oxo, CN, -OR9, -CF3, -OCT, OH2CF3 CF2CF„ -NO2. -NR9R10, N(R9)(CO)R10, N(R9)(CO)OR10, N(R9)(CO)NR9R10, -C( l.)K" (wherein 1. is O or S), -(CO)NR9R10.-O(CO)C,-Cl2alkyl, -O(CO)NR9R10, -COOR9. -SR9. S(O)10R9. SO2NR9R10: SO3H. NH SO2 R9. P(O)R10R12 ; the ring thus formed may further be fused with 1 to 7 membered unsaturated or saturated ring, which may contain from one to three heteroatoins independent!} selected from O. S or N, the fused ring may optionally be substituted with one or more substituents Rc or Rc ;
Rc or Rc is independently selected from the group consisting of but not limited to hydrogen, halogen. C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C1-12 haloalkyl. C2-12 haloalkenyl. C2-12 haloalkynvl. C1-12 alkoxy, C1-12 haloalkoxy, C2,.s cycloalk} I. heterocyclyl. aryl, heteroaryl, (CH2)n-cycloalkyl, (CH2)n-heterocyclyl, (CH2)n-aryl, (CH2)„-heteroaryl. C|-|2 alkylcarbonyl, Cn2 alkoxycarbonyl, CN. OR . -OCF3, -NO2, =NOR10, -NR9R10, N(R9)(CO)RHI. N(R9)(CO)OR'°, N(R9)(CO)NR9R1(I, -C(=L)R9 (wherein L is O or S), -(CO)NR'V1. -C)(CO)R". -O(CO)NR9R10. -COOR9. -SR9, S(O)„,R9. SO2NR9R10; SO3H, NH SO2 R9, P(O)R9R':
R9 and R1 are independently selected from ludrogen. Cn, alkyl, C> L> alkenyl. C2 r alkyinl. C'-,-, ■ haloalkyl, C2-12 haloalkenyl, C1-8 cycloalkyl. heteroc}clyl. aryl. heteroaryl. (CH2)n-cycloalkyl. (CH2)n-heterocyclyl, (CH2)n-aryl, (CH2),-heteroaryl. each of which may be optionally substituted with halogen, hydroxyl or C1-6 alkoxy, or R9and R10 may be joined together to form a heteroevclic or heteroaryl ring which may contain from one to three heteroatoins independently selected from O, S and N, which may optionally be substituted with one or more substituents independent!} selected from Rc or Rc;
m can be 1 or 2;
n can be 1, 2, 3 or 4;
r can be 1, 2, 3 or 4. 2. The compound according to claim 1 having the formula la, wherein
(Formula Removed)
r, Ar, R , R2, R , R , R5, R6, R7 and R8 arc as defined in claim 1; its pharmaceutically accepiabie derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof OR A compound having the Formula lb, wherein
(Formula Removed)
r, Ar, R1, R4, R5, R6, R7 and R's are as defined in claim 1; its pharmaccutically acceptable dematives tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof OR A compound having the Formula Ic wherein
(Formula Removed)
ring A is optionally substituted at any available position by one or more substitucnts independently selected from Rc or Rc ; R2 either represents hvdrogen or a double bond: r. Ar. R . R5 R6. R . R8. R9 and Rc are as defined in claim 1; its pharmaceutically acceptable derivatives, tautomeric lorms. stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof OR A compound having the Formula Id, wherein
(Formula Removed)
X, Y and Z are independently selected from the group consisting of N and -CM: the ring A is optionally substituted at any available position bv one or more siibstituents independently selected from Rc or Rc; r, Ar, R4, R5, R6, R7, R8, Rc and Rc are as defined in claim 1; its pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof OR A compound having the Formula Ie, wherein
(Formula Removed)
ring A is substituted by Rc; r, Ar. Rl, R . R". R . RN and Rc are as defined in claim 1; its
pharmaceutical ly acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs,
metabolites, salts or solvates thereof OR
A compound having the Formula If, wherein
(Formula Removed)
ring A is substituted by Rc or Rc; r, Ar. R1. R5, R1'. R . Rs. Rc and Rc are as defined in claim 1: its pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof OR A compound having the Formula Ig, wherein
(Formula Removed)
ring A is substituted by Rc; r. Ar, R1. R5, R6. R7. R8 and R9 are as defined in claim I: its pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof OR
A compound having the Formula 1h. wherein
(Formula Removed)
ring A is substituted by Rc or Rc; r, Ar, R1. R . R6. R . R8. Rc and Rc are as defined in claim 1: its pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof.
3. The compound according to claim I, wherein Ar is selected from the group consisting of 2,4,5-trifluorophenyl, 2-fluorophenyl. 3.4-difluorophen\ I and 2.5-dilluoropheiiyl.
4. The compound according to claim 1, wherein Rs is selected from the group consisting of hydrogen. fluoro, chloro and methoxy.
5. The compound according to claims 1 and 2. wherein R1 is selected from the group consisting of: hydrogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl. C1-12 haloalkyl, C1-12 haloalkoxy. C2-12 haloalkenvl,
(Formula Removed)
6. The compound according to claims 1 and 2. wherein ring A is substituted with one or more Rc and Rc
independently selected from the group consisting of hydrogen, C1-6 alkyl, C2-12. alkenyl, C2-12 alkynyl,
C3-8cycloalkyl, phenyl, -CH2F, -CHF2, -CF3,, -COOH. -CONH2, -CH2-OCH3, COOC1-6alkyl,
7.
(Formula Removed)
8. The compound according to claim 1 and 2, wherein R1 R2R6 and R7 are hydrogen.
9. A compound which is selected from the group consisting of:
(Formula Removed)
and its pharmaceutically acceptable salts or solv ales thereof. 9. A compound which is selected from the group consisting of:
(Formula Removed)
and its pharmaceutical^ acceptable salts or solvates thereof. 10. A compound which is selected from the group consisting of :
(Formula Removed)
and its pharmaceutically acceptable salts or solvates thereol
1. A compound which is selected from the group consisting of:
(Formula Removed)
and its pharmaceutically acceptable salts or solvates thereof. 12. A compound which is selected from the group consisting of:
(Formula Removed)
wherein r is 1,2,3 or 4
- wherein R8 is selected from a group consisting of H, F, CI and OCH3
wherein R1 is selected from a group consisting of -CH3,
(Formula Removed)
and its pharmaceutically acceptable salt or solvates thererof.
13. A process for the preparation of a compound of formula I. according to claim 1 or its
pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs,
metabolites, salts or solvates thereof, which comprises the following steps:
a) coupling of a compound of Formula II
(Formula Removed)
wherein PG is a protecting group, with a compound of formula III
(Formula Removed)
using coupling conditions, reagents and protecting groups:
b) removing the protecting group (PG) using deprotecling reagent:
wherein r, Ar, R1, R2, R3, R4, R5, R6, R7. and R8 are as defined in claim 1.
14. A pharmaceutical composition, comprising a compound according to claims 1. 8 to 11 or its pharmaceutically acceptable derivatives, tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts or solvates thereof, optionalk in combination with one or more pharmaceulically acceptable carrier(s).
15. Use of a compound according to claims 1. 8 in 11 or its pharmaceutically acceptable derivativcs. tautomeric forms, stereoisomers, polymorphs, prodrugs, metabolites, salts, esters or solvates thereof for the manufacture of a medicament for the prophylaxis, amelioration and/or treatment of one or more condition mediated by DPP-IV in a subject in need thereof, wherein the medicament is administered orally, parenterally or topically.
16. The compounds of Formula I, processes, methods and compositions as described and illustrated herein.