KR20030029061A - Glucagon antagonists/inverse agonists - Google Patents

Abstract

A new class of compounds that act to antagonize the action of glucagon hormones on glucagon receptors. Due to their glucagon receptor antagonistic effects, the compounds may be suitable for the treatment and / or prophylaxis of any disease and condition in which glucagon's antagonistic action is beneficial, such as hyperglycemia, type 1 diabetes, type 2 diabetes, lipid metabolism abnormalities and obesity.

Description

Glucagon antagonists / inverse agonists {GLUCAGON ANTAGONISTS / INVERSE AGONISTS}

Glucagon is a key hormonal agent that regulates the homeostatic regulation of blood glucose levels by interaction with insulin. Glucagon works primarily by stimulating certain cells (mostly hepatocytes) to secrete glucose when blood sugar levels drop. The action of glucagon is the opposite of that of insulin, which stimulates cells to absorb and store glucose every time blood glucose levels rise. Glucagon and insulin are both peptide hormones.

Glucagon is produced in the alpha islets of the pancreas and insulin is produced in the beta islets. Diabetes is a common abnormality in glucose metabolism. The disease may be classified as type 1 diabetes, characterized by hyperglycemia and in insulin-dependent form, or type 2 diabetes, which is non-insulin-dependent in nature. The causes of type 1 diabetes are hyperglycemia and hypoinsulin, and the traditional treatment for this type of disease is the provision of insulin. However, in some diabetic patients of type 1 or type 2, absolute or relatively elevated glucagon levels have been shown to contribute to hyperglycemia. In animal models of type 1 and type 2 diabetes as well as in healthy control animals, removal of circulating glucagon with selective and specific antibodies resulted in a decrease in blood glucose levels (Brand et al., Diabetologia 37, 985 (1994); Diabetes; 43, suppl 1, 172A (1994); Am. J. Physiol. 269, E469-E477 (1995); Diabetes 44 [suppl 1], 134A (1995); Diabetes 45, 1076 (1996). These studies suggest that inhibition of glucagon or antagonizing glucagon may be a useful aid in treating traditional antihyperglycemia in diabetes. The action of glucagon can be inhibited by providing an antagonist or an agonist such as a substance that inhibits or interferes with the glucagon-induced response. Antagonists can naturally be peptides or non-peptides.

Natural glucagon is a 29 amino acid peptide with the following sequence:

His-Ser-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Ser-Arg-Arg-Ala-Gln-Asp-Phe-Val-Gln-Trp- Leu-Met-Asn-Thr-NH ₂ .

Glucagon works by binding to and activating this receptor, which is part of the glucagon-secretin branch of the 7-membrane G-protein coupled to the receptor family (Jelinek et al., Science 259,1614, ( 1993). Receptors function by activating adenylyl cyclase secondary messenger system, resulting in increased cAMP levels.

Many publications disclose peptides that have been described to act as glucagon antagonists. Perhaps the most fully characterized antagonist is DesHis ¹ [Glu ⁹ ] -glucagon amide (Unson et al., Peptides 10,1171 (1989); Post et al., Proc. Natl. Acad. Sci. USA 90,1662) (1993)). Other antagonists include DesHis', Phe ⁶ [Glu ⁹ ] -glucagon amide (Azizh et al., Bioorganic & Medicinal Chem. Lett. 16,1849 (1995)) and NLeu ⁹ , Ala ^11,16 -glucagon amide (Unson et al). , J. Biol. Chem. 269 (17), 12548 (1994).

Peptide antagonists of peptide hormones are often quite potent. However, it is generally known that it is not orally available due to degradation by physiological enzymes and poor diffusion in vivo. Therefore, orally available non-peptide antagonists of peptide hormones are generally preferred. Among non-peptide glucagon antagonists, the quinoxaline derivative, (2-styryl-3- [3- (dimethylamino) propylmethylamino] -6,7-dichloroquinoxaline, has been found to migrate glucagon from rat liver receptors (Collins, JL et al., Bioorganic and Medicinal Chemistry Letters 2 (9): 915-918 (1992)) WO 94/14426, is a natural product of skins comprising pairs of linked 9,10-anthracenedione groups US Pat. No. 4,359,474 discloses glucagon antagonism of 1-phenyl pyrazole derivatives US Pat. No. 4,374,130 discloses substituted disylcyclocyclohexane as a glucagon antagonist. WO 98/04528 to Bayer Corporation discloses substituted pyrimidines and biphenyls as glucagon antagonists US Pat. No. 5,776,954 (Merck & Co., Inc.) discloses substituted pyridyl pyrroles as glucagon antagonists. Ha WO 98/21957, WO 98/22108, WO 98/22109 and US 5,880,139 to Merck & Co., Inc. disclose 2,4-diaryl-5-pyridylimidazoles as glucagon antagonists. 97/16642 and US Pat. No. 5,837,719 to Merck & Co., Inc disclose 2,5-substituted aryl pyrroles as glucagon antagonists WO 98/24780, WO 98/24782, WO 99/24404 and WO 99 / 32448 (Amgen Inc.) discloses substituted pyrimidinone and pyridone compounds and substituted pyrimidine compounds, respectively, which are described as having glucagon antagonistic activity: Madsen et al. (J. Med. Chem. 1998 ( 41) 5151-7) discloses a series of 2- (benzimidazol-2-ylthio) -1- (3,4-dihydroxyphenyl) -1-ethanol as competitive human glucagon receptor antagonists.

WO 99/01423 and WO 00/39088 (Novo Nordisk A / S) disclose different series of alkylidene hydrazides as glucagon antagonists / reverse agonists.

These known glucagon antagonists are structurally different from the present compounds.

Justice

The following is a detailed definition of the terminology used to describe the compounds of the present invention:

"Halogen" refers to an atom selected from the group consisting of F, Cl, Br and I.

As used herein, the term “C _1-6 -alkyl” refers to a saturated, branched or straight chain hydrocarbon having 1 to 6 carbon atoms. Representative examples include methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, isohexyl, etc. But not limited to this.

As used herein, the term “C _2-6 -alkenyl” refers to a branched or straight chain hydrocarbon group having 2 to 6 carbon atoms and at least one double bond. Examples of this group include vinyl, 1-propenyl, 2-propenyl, iso-propenyl, 1,3-butadienyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-1 -Propenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 3-methyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 2, 4-hexadienyl, 5-hexenyl, and the like, but is not limited thereto.

As used herein, the term "C _2-6 -alkynyl" refers to a group of branched or straight chain hydrocarbons having 2 to 6 carbon atoms and at least one triple bond. Examples of this group are ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4- Pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, 2,4helsadininyl, and the like.

As used herein, the term “C _1-6 -alkoxy” refers to the radical —OC _1-6 -alkyl, wherein C _1-6 -alkyl is as defined above. Representative examples include methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, sec-butoxy, tert-butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy and the like.

As used herein, the term “C _1-6 -alkanoyl” means a —C (O) H or —C (O) —C _1-5 -alkyl group. Representative examples are formyl, acetyl, propionyl, butyryl, valeryl, hexanoyl and the like.

The term "C _3-8 -cycloalkyl" as used herein refers to a saturated carbocyclic group having 3 to 8 carbon atoms. Representative examples are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.

The term "C _{4-8 -cycloalkenyl} " as used herein denotes a non-aromatic carbocyclic group having 4 to 8 carbon atoms comprising one or two double bonds. Representative examples include 1-cyclopentenyl, 2-cyclopentenyl, 3-cyclopentenyl, 1-cyclohexenyl, 2-cyclohexenyl, 3-cyclohexenyl, 2-cycloheptenyl, 3-cycloheptenyl , 2-cyclooctenyl, 1,4-cyclooctadienyl and the like.

As used herein, the term “heterocyclyl” refers to a non-aromatic 3 to 10 ring comprising one or more heteroatoms selected from nitrogen, oxygen and sulfur and optionally one or two double bonds. Representative examples are pyrrolidinyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl, aziridinyl, tetrahydrofuranyl and the like.

As used herein, the term “aryl” is intended to include carbocyclic aromatic ring systems such as phenyl, biphenylyl, naphthyl, anthracenyl, phenanthrenyl, fluorenine, indenyl, pentalenyl, azulenin, and the like. Aryl is also intended to include partially hydrogenated derivatives of the carbocyclic systems listed above. Non-limiting examples of such partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthyl, 1,4-dihydronaphthyl and the like.

As used herein, the term “arylene” refers to a divalent carbon ring aromatic ring system such as phenylene, biphenylylene, naphthylene, anthracenylene, phenanthrenylene, fluorenylene, indenylene, pentalenylene, azulenylene, and the like. It is intended to include. Arylene is also intended to include partially hydrogenated derivatives of the above listed carbocyclic systems. Non-limiting examples of such partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthylene, 1,4-dihydronaphthylene and the like.

As used herein, the term "aryloxy" refers to an -O-aryl group, where aryl is as defined above.

As used herein, the term “aroyl” refers to a —C (O) -aryl group, where aryl is as defined above.

The term "heteroaryl" as used herein refers to furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4 -Triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,4-triazinyl, 1, 2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1, 2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, tetrazolyl, thiadiazinyl, indolyl, isoindoleyl, benzofuryl, benzothienin, indah Zolyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, purinyl, quinazolinyl, quinolinyl, quinolinyl, isoquinolinyl, quinoxalinyl, naphthyridinyl Nitrogen, such as neil, putridinyl, carbazolyl, azepinyl, diazepinyl, acridinyl, etc. It is intended to include heterocyclic aromatic ring systems containing one or more heteroatoms selected from cattle and sulfur. Heteroaryl is also intended to include partially hydrogenated derivatives of the heterocyclic systems listed above. Non-limiting examples of such partially hydrogenated derivatives are 2,3-dihydrobenzofuranyl, pyrrolinyl, pyrazolinyl, indolinyl, oxazolidinyl, oxazolinyl, oxazinyl, and the like.

“Aryl-C _1-6 -alkyl”, “heteroaryl-C _1-6 -alkyl”, “aryl-C _2-6 -alkenyl”, and the like, are substituted above with aryl or heteroaryl as defined above. C _1-6 -alkyl or C _1-6 -alkenyl as defined. For example:

As used herein, the term "optionally substituted" means that the group in question is unsubstituted or substituted with one or more of the listed substituents. The substituents may be the same or different when the group in question is substituted with one or more substituents.

Any of the terms defined above may appear more than once in the structural formula, and after this occurrence each term may be defined independently and differently.

In addition, it should be understood that the groups in question may be the same or different when using the terms "independently" and "independently selected from."

The present invention relates to agents that act to counteract the action of glucagon peptide hormones on glucagon receptors. More particularly, it relates to glucagon antagonists or inverse agonists.

The present invention is based on the unexpected observation that the compounds of formula (I) disclosed below antagonize the action of glucagon.

Compounds are advantageous by being selective towards glucagon receptors and exhibit higher binding affinity for glucagon receptors as compared to binding affinity for structurally related GIP (digestion inhibiting peptide) receptors and GLP-1 receptors.

Accordingly, the present invention relates to compounds of formula (I) as well as to any optical or geometric isomers or tautomers or mixtures thereof or pharmaceutically acceptable salts thereof:

In the above formula

R ¹ , R ² , R ³ , R ⁴ and R ⁵ are independently hydrogen or C _1-6 -alkyl,

A is -C (O)-, -CH (OR⁶)- Or -CHF-,

Wherein R ⁶ is hydrogen or C _1-6 -alkyl,

Z is a divalent radical derived from arylene or a 5 or 6 membered heteroaromatic ring containing 1 or 2 heteroatoms selected from nitrogen, oxygen and sulfur,

This _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 9, -NR 9 R 10 and C _1-6 - optionally with one or two groups selected from alkyl R ⁷ and R ⁸ Can be substituted,

Wherein R ⁹ and R ¹⁰ are independently hydrogen or C _1-6 -alkyl,

X is

But here

r is 0 or 1,

q and s are independently 0, 1, 2 or 3,

R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are independently hydrogen, C _1-6 -alkyl or C _3-8 -cycloalkyl,

D is

But here

R ¹⁵ , R ¹⁶ R ¹⁷ and R ¹⁸ are independently;

Hydrogen, halogen, -CN, -CHF ₂ , -CF ₃ , -OCF ₃ , -OCHF ₂ , -OCH ₂ CF ₃ , -OCF ₂ CHF ₂ , -S (O) ₂ CF ₃ , -SCF ₃ ,- NO ₂ , -OR ²¹ , -NR ²¹ R ²² , -SR ²¹ , -NR ²¹ S (O) ₂ R ²² , -S (O) ₂ NR ²¹ R ²² , -S (O) NR ²¹ R ²² ,- S (O) R ²¹ , -S (O) ₂ R ²¹ , -C (O) NR ²¹ R ²² , -OC (O) NR ²¹ R ²² , -NR ²¹ C (O) R ²² , -CH ₂ C (O) NR ²¹ R ²² , -OCH ₂ C (O) NR ²¹ R ²² , -OC (O) R ²¹ , -C (O) R ²¹ or -C (O) OR ²¹ ,

C _1-6 -alkyl, C _2-6 -alkenyl or C _2-6 -alkynyl,

(Which is optionally _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 21, -NR 21 R 22 and C _1-6 - may be substituted with one or more substituents selected from alkyl)

C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl, heterocyclyl, C _3-8 -cycloalkyl-C _1-6 -alkyl, C _3-8 -cycloalkyl-C _1-6- Alkoxy, C _3-8 -cycloalkyloxy, C _3-8 -cycloalkyl-C _1-6 -alkylthio, C _3-8 -cycloalkylthio, C _3-8 -cycloalkyl-C _2-6 -al alkenyl, C _3-8 - cycloalkyl, -C _2-6 - alkynyl, C _4-8 - cycloalkenyl -C _1-6 - alkyl, C _4-8 - cycloalkenyl -C _2-6 - alkenyl , C _4-8 - cycloalkenyl -C _2-6 - alkynyl, heterocyclyl, -C _1-6 - alkyl, heterocyclyl, -C _2-6 - alkenyl, heterocyclyl, -C _2-6 - Alkynyl, aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Ci _-6 -alkoxy, aryl-C _1-6 -alkyl, aryl-C _2-6 -alkenyl, aryl-C _2-6 -Alkynyl, heteroaryl, heteroaryl-C _1-6 -alkyl, heteroaryl-C _2-6 -alkenyl or heteroaryl-C _2-6 -alkynyl,

Wherein the ring portion is optionally selected from halogen, —C (O) OR ²¹ , —CN, —CF ₃ , —OCF ₃ , —N0 ₂ , —OR ²¹ , —NR ²¹ R ²² and C _1-6 -alkyl May be substituted with one or more substituents,

Wherein R ²¹ and R ²² are independently hydrogen, C _1-6 -alkyl, aryl-C _1-6 -alkyl or aryl,

Or R ²¹ and R ²² optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally comprise one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; Can form a 3 to 8 membered heterocyclic ring,

Or when two of the groups R ¹⁵ to R ¹⁸ are located together at adjacent points, they may form a crosslinking-(CR ²³ R ²⁴ ) _a -O- (CR ²⁵ R ²⁶ ) _c -O-

From here

a is 0, 1 or 2,

c is 1 or 2,

R ²³ , R ²⁴ , R ²⁵ and R ²⁶ are independently hydrogen, C _1-6 -alkyl or fluorine,

R ¹⁹ and R ²⁰ are independently hydrogen, C _1-6 -alkyl, C _3-8 -cycloalkyl or C _3-8 -cycloalkyl-C _1-6 -alkyl,

E is

But here

R ²⁷ and R ²⁸ are independently

Hydrogen, halogen, -CN, -CF ₃ , -OR ³² , -NR ³² R ³³ , C _1-6 -alkyl, C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl or aryl,

Wherein the aryl group may be optionally substituted with one or more substituents selected from halogen, —CN, —CF ₃ , —NO ₂ , —OR ³² , —NR ³² R ³³ and C _1-6 -alkyl,

Wherein R ³² and R ³³ are independently hydrogen or C _1-6 -alkyl,

R ³² and R ³³ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally comprise one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; To form a 3 to 8 membered heterocyclic ring,

R ²⁹ , R ³⁰ and R ³¹ are independently;

Hydrogen, halogen, -CHF ₂ , -CF ₃ , -OCF ₃ , -OCHF ₂ , -OCH ₂ CF ₃ , -OCF ₂ CHF ₂ , -SCF ₃ , -OR ³⁴ , -NR ³⁴ R ³⁵ , -SR ³⁴ , -S (O) R ³⁴ , -S (O) ₂ R ³⁴ , -C (O) NR ³⁴ R ³⁵ , -OC (O) NR ³⁴ R ³⁵ , -NR ³⁴ C (O) R ³⁵ , -OCH ₂ C (O) NR ³⁴ R ³⁵ , -C (O) R ³⁴ or -C (O) OR ³⁴ ,

C _1-6 -alkyl, C _2-6 -alkenyl or C _2-6 -alkynyl,

(It may be optionally substituted with one or more substituents selected from halogen, —CN, —CF ₃ , —OCF ₃ , —N0 ₂ , —OR ³⁴ , —NR ³⁴ R ³⁵ and C _1-6 -alkyl)

C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl, heterocyclyl, C _3-8 -cycloalkyl-C _1-6 -alkyl, C _3-8 -cycloalkyl-C _2-6- alkenyl, C _3-8 - cycloalkyl, -C _2-6 - alkynyl, C _4-8 - cycloalkenyl -C _1-6 - alkyl, C _4-8 - cycloalkenyl -C _2-6 - Al alkenyl, C _4-8 - cycloalkenyl -C _2-6 - alkynyl, heterocyclyl, -C _1-6 - alkyl, heterocyclyl, -C _2-6 - alkenyl, heterocyclyl, -C _2-6 -Alkynyl, aryl, aryloxy, aroyl, aryl-C _1-6 -alkoxy, aryl-C _1-6 -alkyl, aryl-C _2-6 -alkenyl, aryl-C _2-6 -alkynyl, Heteroaryl, heteroaryl-C _1-6 -alkyl, heteroaryl-C _2-6 -alkenyl or heteroaryl-C _2-6 -alkynyl,

Wherein the ring moiety is halogen, -CN, -CF ₃ , -OCF ₃ , -NO ₂ , -OR ³⁴ , -NR ³⁴ R ³⁵ and

Optionally substituted with one or more substituents selected from C _1-6 -alkyl,

Wherein R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl,

Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur when optionally attached to the same nitrogen atom together with said nitrogen atom and optionally comprise one or two double bonds; To form a 3 to 8 membered heterocyclic ring containing,

Or the radicals —O— (CH ₂ ) _t —CR ³⁶ R ³⁷ — (CH ₂ ) ₁ —O when two of the groups R ²⁹ , R ³⁰ and R ³¹ are attached together to a carbon atom of the same ring or to a carbon atom of a different ring _{-, - (CH 2) t} -CR 36 R 37 - (CH 2) l - , or ^{_{-S- (CH 2) t -CR 36}} R 37 - (CH 2) l can form a -S-,

From here

t and l are independently 0,1,2,3,4 or 5,

R ³⁶ and R ³⁷ are independently hydrogen or C _1-6 -alkyl.

In another aspect, the invention relates to compounds of formula (I ') as well as to any optical or geometric isomers or tautomers or mixtures thereof or pharmaceutically acceptable salts thereof:

Formula I '

In the above formula

R ¹ , R ² , R ³ , R ⁴ and R ⁵ are independently hydrogen or C _1-6 -alkyl,

A is -C (O)-, -CH (OR ⁶ )-or -CHF-,

Where R ⁶ is hydrogen, C _1-6 -alkyl,

Z is a divalent radical derived from arylene or a 5 or 6 membered heteroaromatic ring containing 1 or 2 heteroatoms selected from nitrogen, oxygen and sulfur,

This _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 9, -NR 9 R 10 and C _1-6 - alkyl optionally substituted by one or two groups selected from R ⁷ and R ⁸ Wherein R ⁹ and R ¹⁰ are independently hydrogen or C _1-6 -alkyl,

X is

But here

r is 0 or 1,

q and s are independently 0, 1, 2 or 3,

R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are independently hydrogen, C _1-6 -alkyl or C _3-8 -cycloalkyl,

D is

But here

R ¹⁵ , R ¹⁶ R ¹⁷ and R ¹³ are independently;

Hydrogen, halogen, -CN, -CH ₂ CN, -CHF ₂ , -CF ₃ , -OCF ₃ , -OCHF ₂ , -OCH ₂ CF ₃ , -OCF ₂ CHF ₂ , -S (O) ₂ CF ₃ , -SCF ₃ , -NO ₂ , -OR ²¹ , -NR ²¹ R ²² , -SR ²¹ , -NR ²¹ S (O) ₂ R ²² , -S (O) ₂ NR ²¹ R ²² , -S (O) NR ²¹ R ²² , -S (O) R ²¹ , -S (O) ₂ R ²¹ , -C (O) NR ²¹ R ²² , -OC (O) NR ²¹ R ²² , -NR ²¹ C (O) R ²² , -CH ₂ C (O) NR ²¹ R ²² , -OCH ₂ C (O) NR ²¹ R ²² , -CH ₂ OR ²¹ , -CH ₂ NR ²¹ R ²² , -OC (O) R ²¹ , -C ( O) R ²¹ or -C (O) OR ²¹ ,

C _1-6 -alkyl, C _2-6 -alkenyl or C _2-6 -alkynyl,

(Which is optionally _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 21, -NR 21 R 22 and C _1-6 - may be substituted with one or more substituents selected from alkyl)

C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl, heterocyclyl, C _3-8 -cycloalkyl-C _1-6 -alkyl, C _3-8 -cycloalkyl-C _1-6- Alkoxy, C _3-8 -cycloalkyloxy, C _3-8 -cycloalkyl-C _1-6 -alkylthio, C _3-8 -cycloalkylthio, C _3-8 -cycloalkyl-C _2-6 -al alkenyl, C _3-8 - cycloalkyl, -C _2-6 - alkynyl, C _4-8 - cycloalkenyl -C _1-6 - alkyl, C _4-8 - cycloalkenyl -C _2-6 - alkenyl , C _4-8 - cycloalkenyl -C _2-6 - alkynyl, heterocyclyl, -C _1-6 - alkyl, heterocyclyl, -C _2-6 - alkenyl, heterocyclyl, -C _2-6 - Alkynyl, aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Ci _-6 -alkoxy, aryl-C _1-6 -alkyl, aryl-C _2-6 -alkenyl, aryl-C _2-6 -Alkynyl, heteroaryl, heteroaryl-C _1-6 -alkyl, heteroaryl-C _2-6 -alkenyl or heteroaryl-C _2-6 -alkynyl,

Wherein the ring portion is optionally selected from halogen, -C (O) OR, -CN, -CF ₃ , -OCF ₃ , -N0 ₂ , -OR ²¹ , -NR ²¹ R ²² and C _1-6 -alkyl May be substituted with more than one substituent,

Wherein R ²¹ and R ²² are independently hydrogen, C _1-6 -alkyl, aryl-C _1-6 -alkyl or aryl,

Or R ²¹ and R ²² optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally comprise one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; Can form a 3 to 8 membered heterocyclic ring,

Or when two of the groups R ¹⁵ to R ¹⁸ are located together at adjacent points, they may form a crosslinking-(CR ²³ R ²⁴ ) _a -O- (CR ²⁵ R ²⁶ ) _c -O-

From here

a is 0, 1 or 2,

c is 1 or 2,

R ²³ , R ²⁴ , R ²⁵ and R ²⁶ are independently hydrogen, C _1-6 -alkyl or fluorine,

R ¹⁹ and R ²⁰ are independently hydrogen, C _1-6 -alkyl, C _3-8 -cycloalkyl or C _3-8 -cycloalkyl-C _1-6 -alkyl,

E is

But here

R ²⁷ and R ²³ are independently

Hydrogen, halogen, -CN, -CF ₃ , -OR ³² , -NR ³² R ³³ , C _1-6 -alkyl, C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl or aryl,

Wherein the aryl group may be optionally substituted with one or more substituents selected from halogen, —CN, —CF ₃ , —NO ₂ , —OR ³² , —NR ³² R ³³ and C _1-6 -alkyl,

Wherein R ³² and R ³³ are independently hydrogen or C _1-6 -alkyl,

R ³² and R ³³ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally comprise one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; To form a 3 to 8 membered heterocyclic ring,

R ²⁹ , R ³⁰ and R ³¹ are independently;

Hydrogen, halogen, -CHF ₂ , -CF ₃ , -OCF ₃ , -OCHF ₂ , -OCH ₂ CF ₃ , -OCF ₂ CHF ₂ , -SCF ₃ , -OR ³⁴ , -NR ³⁴ R ³⁵ , -SR ³⁴ , -S (O) R ³⁴ , -S (O) ₂ R ³⁴ , -C (O) NR ³⁴ R ³⁵ , -OC (O) NR ³⁴ R ³⁵ , -NR ³⁴ C (O) R ³⁵ , -OCH ₂ C (O) NR ³⁴ R ³⁵ , -C (O) R ³⁴ or -C (O) OR ³⁴ ,

C _1-6 -alkyl, C _2-6 -alkenyl or C _2-6 -alkynyl,

(It may be optionally substituted with one or more substituents selected from halogen, —CN, —CF ₃ , —OCF ₃ , —N0 ₂ , —OR ³⁴ , —NR ³⁴ R ³⁵ and C _1-6 -alkyl)

C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl, heterocyclyl, C _3-8 -cycloalkyl-C _1-6 -alkyl, C _3-8 -cycloalkyl-C _2-6- alkenyl, C _3-8 - cycloalkyl, -C _2-6 - alkynyl, C _4-8 - cycloalkenyl -C _1-6 - alkyl, C _4-8 - cycloalkenyl -C _2-6 - Al alkenyl, C _4-8 - cycloalkenyl -C _2-6 - alkynyl, heterocyclyl, -C _1-6 - alkyl, heterocyclyl, -C _2-6 - alkenyl, heterocyclyl, -C _2-6 -Alkynyl, aryl, aryloxy, aroyl, aryl-C _1-6 -alkoxy, aryl-C _1-6 -alkyl, aryl-C _2-6 -alkenyl, aryl-C _2-6 -alkynyl, Heteroaryl, heteroaryl-C _1-6 -alkyl, heteroaryl-C _2-6 -alkenyl or heteroaryl-C _2-6 -alkynyl,

Wherein the ring moiety is halogen, -CN, -CF ₃ , -OCF ₃ , -NO ₂ , -OR ³⁴ , -NR ³⁴ R ³⁵ and

Optionally substituted with one or more substituents selected from C _1-6 -alkyl,

Wherein R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl,

Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur when optionally attached to the same nitrogen atom together with said nitrogen atom and optionally comprise one or two double bonds; To form a 3 to 8 membered heterocyclic ring containing,

Or the radicals —O— (CH ₂ ) _t —CR ³⁶ R ³⁷ — (CH ₂ ) ₁ —O when two of the groups R ²⁹ , R ³⁰ and R ³¹ are attached together to a carbon atom of the same ring or to a carbon atom of a different ring _{-, - (CH 2) t} -CR 36 R 37 - (CH 2) l - , or ^{_{-S- (CH 2) t -CR 36}} R 37 - (CH 2) l can form a -S-,

From here

t and l are independently 0,1,2,3,4 or 5,

R ³⁶ and R ³⁷ are independently hydrogen or C _1-6 -alkyl.

In another embodiment, the invention relates to any optical or geometric isomer or tautomer or mixture thereof or pharmaceutically acceptable salt thereof, as well as to a compound of formula (I):

Formula I "

Wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are independently hydrogen or C _1-6 -alkyl,

A is -C (O)-, -CH (OR⁶)- Or -CHF-,

Wherein R ⁶ is hydrogen or C _1-6 -alkyl,

Z is a divalent radical derived from arylene or a 5 or 6 membered heteroaromatic ring containing 1 or 2 heteroatoms selected from nitrogen, oxygen and sulfur,

This _{halogen, -CN, CF 3, -OCF 3} , -NO 2, -OR 9, -NR 9 R 10 and C _1-6 - alkyl, or one selected from the two groups to be optionally substituted with R ⁷ and R ⁸ Wherein R ⁹ and R ¹⁰ are independently hydrogen or C _1-6 -alkyl,

X is

But here

r is 0 or 1,

q and s are independently 0, 1, 2 or 3,

R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are independently hydrogen or C _1-6 -alkyl,

D is

But here

R ¹⁵ , R ¹⁶ R ¹⁷ and R ¹³ are independently;

Hydrogen, halogen, -CN, -CN ₂ CN, -CHF ₂ , -CF ₃ , -OCF ₃ , -OCHF ₂ , -OCH ₂ CF ₃ , -OCF ₂ CHF ₂ , -OS (O) ₂ CF ₃ , -SCF ₃ , -NO ₂ , -OR ²¹ , -NR ²¹ R ²² , -SR ²¹ , -NR ²¹ S (O) ₂ R ²² , -S (O) ₂ NR ²¹ R ²² , -S (O) NR ²¹ R ²² , -S (O) R ²¹ , -S (O) ₂ R ²¹ , -OS (O) 2R ²¹ , -C (O) NR ²¹ R ²² , -OC (O) NR ²¹ R ²² ,- NR ²¹ C (O) R ²² , -CH ₂ C (O) NR ²¹ R ²² , -OCH ₂ C (O) NR ²¹ R ²² , -CH ₂ OR ²¹ , -CH ₂ NR ²¹ R ²² , -OC ( O) R ²¹ , -C (O) R ²¹ or -C (O) OR ²¹ ,

C _1-6 -alkyl, C _2-6 -alkenyl or C _2-6 -alkynyl,

(Which is optionally _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 21, -NR 21 R 22 and C _1-6 - may be substituted with one or more substituents selected from alkyl)

C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl, heterocyclyl, C _3-8 -cycloalkyl-C _1-6 -alkyl, C _3-8 -cycloalkyl-C _1-6- Alkoxy, C _3-8 -cycloalkyloxy, C _3-8 -cycloalkyl-C _1-6 -alkylthio, C _3-8 -cycloalkylthio, C _3-8 -cycloalkyl-C _2-6 -al alkenyl, C _3-8 - cycloalkyl, -C _2-6 - alkynyl, C _4-8 - cycloalkenyl -C _1-6 - alkyl, C _4-8 - cycloalkenyl -C _2-6 - alkenyl , C _4-8 - cycloalkenyl -C _2-6 - alkynyl, heterocyclyl, -C _1-6 - alkyl, heterocyclyl, -C _2-6 - alkenyl, heterocyclyl, -C _2-6 - Alkynyl, aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Ci _-6 -alkoxy, aryl-C _1-6 -alkyl, aryl-C _2-6 -alkenyl, aryl-C _2-6 -Alkynyl, heteroaryl, heteroaryl-C _1-6 -alkyl, heteroaryl-C _2-6 -alkenyl or heteroaryl-C _2-6 -alkynyl,

Wherein the ring moiety may be optionally substituted with one or more substituents selected from halogen, —CN, —CF ₃ , —OCF ₃ , —N0 ₂ , —OR ²¹ , —NR ²¹ R ²² and C _1-6 -alkyl; ,

Wherein R ²¹ and R ²² are independently hydrogen, C _1-6 -alkyl or aryl, or R ²¹ and R ²² are selected from nitrogen, oxygen and sulfur when attached to the same nitrogen atom together with said nitrogen atom Or may further form a 3-8 membered heterocyclic ring optionally further comprising two heteroatoms and optionally comprising one or two double bonds,

Or when two of the groups R ¹⁵ to R ¹⁸ are located together at adjacent points, they may form a crosslinking-(CR ²³ R ²⁴ ) _a -O- (CR ²⁵ R ²⁶ ) _c -O-

From here

a is 0, 1 or 2,

c is 1 or 2,

R ²³ , R ²⁴ , R ²⁵ and R ²⁶ are independently hydrogen, C _1-6 -alkyl or fluorine,

R ¹⁹ and R ²⁰ are independently hydrogen, C _1-6 -alkyl, C _3-8 -cycloalkyl or C _3-8 -cycloalkyl-C _1-6 -alkyl,

E is

But here

R ²⁷ and R ²⁸ are independently

Hydrogen, halogen, -CN, -CF ₃ , -OR ³² , -NR ³² R ³³ , C _1-6 -alkyl, C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl or aryl,

Wherein the aryl group may be optionally substituted with one or more substituents selected from halogen, —CN, —CF ₃ , —NO ₂ , —OR ³² , —NR ³² R ³³ and C _1-6 -alkyl,

Wherein R ³² and R ³³ are independently hydrogen or C _1-6 -alkyl,

R ³² and R ³³ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally comprise one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; To form a 3 to 8 membered heterocyclic ring,

R ²⁹ , R ³⁰ and R ³¹ are independently;

And _{hydrogen, -CHF 2, -CF 3, -OCF} 3, -OCHF 2, -OCH 2 CF 3, -OCF 2 CHF 2, -SCF 3, -OR 34, -NR 34 R 35, -SR 34, - S (O) R ³⁴ , -S (O) ₂ R ³⁴ , -C (O) NR ³⁴ R ³⁵ , -OC (O) NR ³⁴ R ³⁵ , -NR ³⁴ C (O) R ³⁵ , -OCH ₂ C (O) NR ³⁴ R ³⁵ , -C (O) R ³⁴ or -C (O) OR ³⁴ ,

C _1-6 -alkyl, C _2-6 -alkenyl or C _2-6 -alkynyl,

(It may be optionally substituted with one or more substituents selected from halogen, —CN, —CF ₃ , —OCF ₃ , —N0 ₂ , —OR ³⁴ , —NR ³⁴ R ³⁵ and C _1-6 -alkyl)

C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl, heterocyclyl, C _3-8 -cycloalkyl-C _1-6 -alkyl, C _3-8 -cycloalkyl-C _2-6- alkenyl, C _3-8 - cycloalkyl, -C _2-6 - alkynyl, C _4-8 - cycloalkenyl -C _1-6 - alkyl, C _4-8 - cycloalkenyl -C _2-6 - Al alkenyl, C _4-8 - cycloalkenyl -C _2-6 - alkynyl, heterocyclyl, -C _1-6 - alkyl, heterocyclyl, -C _2-6 - alkenyl, heterocyclyl, -C _2-6 -Alkynyl, aryl, aryloxy, aroyl, aryl-C _1-6 -alkoxy, aryl-C _1-6 -alkyl, aryl-C _2-6 -alkenyl, aryl-C _2-6 -alkynyl, Heteroaryl, heteroaryl-C _1-6 -alkyl, heteroaryl-C _2-6 -alkenyl or heteroaryl-C _2-6 -alkynyl,

Wherein the ring moiety is halogen, -CN, -CF ₃ , -OCF ₃ , -NO ₂ , -OR ³⁴ , -NR ³⁴ R ³⁵ and

Optionally substituted with one or more substituents selected from C _1-6 -alkyl,

Wherein R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl,

R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally comprise one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; To form a 3 to 8 membered heterocyclic ring,

Or the radicals —O— (CH ₂ ) _t —CR ³⁶ R ³⁷ — (CH ₂ ) ₁ —O when two of the groups R ²⁹ , R ³⁰ and R ³¹ are attached together to a carbon atom of the same ring or to a carbon atom of a different ring _{-, - (CH 2) t} -CR 36 R 37 - (CH 2) l - , or ^{_{-S- (CH 2) t -CR 36}} R 37 - (CH 2) l can form a -S-,

From here

t and l are independently 0,1,2,3,4 or 5,

R ³⁶ and R ³⁷ are independently hydrogen or C _1-6 -alkyl.

In one embodiment, R ¹ , R ² , R ³ , R ⁴ and R ⁵ are hydrogen.

In one embodiment, A is -CHF-.

In another embodiment, A is -CH (OR ⁶ )-, wherein R ⁶ is as defined in formula (I), such as -CH (OH)-.

In one embodiment, Z is

Where R ⁷ and R ⁸ are

As defined for Formula (I).

In one embodiment, X is

Wherein q is 0 or 1, r is 0 or 1, s is 0, 1 or 2, and R ¹² and R ¹³ are independently hydrogen or C _1-6 -alkyl.

In another embodiment, X is -C (O) NH-, -C (O) NHCH _2- , -C (O) NHCH (CH ₃ )-, -C (O) NHC (CH ₃ ) _2- , -C (O) NHCH ₂ CH _2- , -C (O) CH _2- , C (O) CH ₂ CH _2- , -C (O) CH = CH-,-(CH ₂ ) _s- , -C (O)-, -C (O) O- or -NHC (O)-, where s is 0 or 1.

In yet another embodiment, X is -C (O) NH-, -C (O) NHCH _2- , -C (O) NHCH (CH ₃ )-, -C (O) NHCH ₂ CH ₂ -,- C (O) CH _2- , -C (O) CH = CH-,-(CH ₂ ) _s- , -C (O)-, -C (O) O- or -NHC (O)-, wherein Where s is 0 or 1.

In yet another embodiment, X is -C (O) NH-, -C (O) NHCH _2- , -C (O) NHCH (CH ₃ )-, -C (O) NHCH ₂ CH ₂ -,- C (O) CH ₂ —, —CH ₂ —, —C (O) — or —NHC (O) —.

In still further embodiments, X is -C (O) NH-, -C (O) NHCH _2- , -C (O) NHCH (CH ₃ )-, -C (O) CH ₂ -or -C ( O)-, for example -C (O) NH-.

In one embodiment, D is

Wherein R ¹⁵ , R ¹⁶ , R ¹⁷ , R ¹⁸ , R ¹⁹ and R ²⁰ are as defined for Formula (I).

In another embodiment, D is

Wherein R ¹⁵ , R ¹⁶ and R ¹⁷ are as defined for Formula (I).

In one embodiment, R ¹⁵ , R ¹⁶ and R ¹⁷ are independently hydrogen, halogen, -CN, -NO ₂ , -CF ₃ , -OCF ₃ , -SCF ₃ , C _1-6 -alkyl, C _1-6 -Alkoxy, -SC _1-6 -alkyl, -C (O) OR ²¹ , -C (O) R ²¹ , -CH ₂ OR ²¹ , -C (O) NR ²¹ R ²² , -S (O) R ²¹ , -S (O) ₂ R ²¹ , -S (O) ₂ CF ₃ , -S (O) ₂ NR ²¹ R ²² , C _3-8 -cycloalkyl, C _3-8 -cycloalkyl-C _{1-6 -Alkoxy} or C _3-8 -cycloalkyl-C _1-6 -thioalkyl, or

Aryl, heteroaryl or aryloxy, which may be optionally substituted by -CF ₃ , -OCF ₃ , C _1-6 -alkyl, halogen or -C (O) OR ²¹ , or

When two of R ¹⁵ , R ¹⁶ and R ¹⁷ are located together at adjacent points they form a crosslinking-(CR ²³ R ²⁴ ) _a -O- (CR ²⁵ R ²⁶ ) _c -O-,

Wherein R ²¹ and R ²² are independently hydrogen or C _1-6 -alkyl and a, c, R ²³ , R ²⁴ , R ²⁵ and R ²⁶ are as defined for formula (I).

In another embodiment, R ¹⁵ , R ¹⁶ and R ¹⁷ are independently hydrogen, halogen, -CN, -CF ₃ , -OCF ₃ or C _1-6 -alkoxy or R ¹⁵ , R ¹⁶ crosslinked together -CF Forms ₂ -O-CF ₂ -O- and R ¹⁷ is hydrogen.

In another embodiment, R ¹⁵ , R ¹⁶ and R ¹⁷ are independently hydrogen, halogen, -CN, -CF ₃ , -OCF ₃ or C _1-6 -alkyl.

In another embodiment, D is

Wherein R ¹⁵ , R ¹⁶ , R ¹⁹ and R ²⁰ are as defined for Formula (I).

In yet another embodiment, D is

Wherein R ¹⁵ and R ¹⁶ are both hydrogen and R ¹⁹ is C _1-6 -alkyl, C _3-8 -cycloalkyl or C _3-8 -cycloalkyl-C _1-6 -alkyl.

In yet another embodiment, D is

Wherein R ¹⁵ and R ¹⁶ are both hydrogen and R ¹⁹ and R ²⁰ are both C _1-6 -alkyl.

In one embodiment, E is

Wherein R ²⁷ , R ²⁸ , R ²⁹ , R ³⁰ and R ³¹ are as defined for Formula (I).

In another embodiment, E is

Wherein R ²⁷ and R ²⁸ are as defined for Formula (I).

In yet another embodiment, E is

Wherein R ²⁷ and R ²⁸ are as defined for Formula (I).

In one embodiment, R ²⁷ and R ²⁸ are independently hydrogen, C _1-6 -alkyl, C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl or phenyl, wherein the phenyl group is Optionally substituted as defined.

In another embodiment, R ²⁷ and R ²⁸ are independently hydrogen, C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl.

In yet another embodiment, R ²⁷ is hydrogen and R ²⁸ is C _1-6 -alkyl or C _3-8 -cycloalkyl, for example tert-butyl, cyclohexyl or cyclohexenyl.

In another embodiment, E is

Wherein R ²⁹ , R ³⁰ and R ³¹ are as defined for Formula (I).

In yet another embodiment, E

Wherein R ²⁹ , R ³⁰ and R ³¹ are as defined for Formula (I).

In one embodiment, R ²⁹ , R ³⁰ and R ³¹ are independently

And _{hydrogen, -CHF 2, -CF 3, -OCF} 3, -OCHF 2, -OCH 2 CF 3, -OCF 2 CHF 2, -SCF 3, -OR 34, -NR 34 R 35, -SR 34, - S (O) R ³⁴ , -S (O) ₂ R ³⁴ , -C (O) NR ³⁴ R ³⁵ , -OC (O) NR ³⁴ R ³⁵ , -NR ³⁴ C (O) R ³⁵ , -OCH ₂ C (O) NR ³⁴ R ³⁵ , -C (O) R ³⁴ or -C (O) OR ³⁴ ,

C _1-6 -ylkyl, C _2-6 -alkenyl or C _2-6 -alkynyl,

(Which is a _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - may be optionally substituted with one or more substituents selected from alkyl)

C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl,

This _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - can be optionally substituted with one or more substituents selected from alkyl,

Wherein R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl,

Or when R ³⁴ and R ³⁵ are attached together with the nitrogen atom to the same nitrogen atom, optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally comprise one or two double bonds To form a 3 to 8 membered heterocyclic ring.

In yet another embodiment, R ²⁹ , R ³⁰ and R ³¹ are independently

Hydrogen, C _1-6 -alkoxy, halogen, —CF ₃ , —OCF ₃ or R ³⁴ and R ³⁵ are —NR ³⁴ R ³⁵ as defined for Formula (I), or

C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl, optionally substituted as defined for Formula (I).

In yet another embodiment, R ²⁹ , R ³⁰ and R ³¹ are independently

Hydrogen or

C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl, optionally substituted as defined for Formula (I).

In yet another embodiment, R ²⁹ , R ³⁰ and R ³¹ are independently

Hydrogen or

C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl,

And which _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - can be optionally substituted with one or more substituents selected from alkyl,

Where R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl,

Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; It can form a 3 to 8 membered heterocyclic ring including.

In another embodiment, R ²⁹ and R ³¹ are both hydrogen and R ³⁰ is different from hydrogen.

In yet another embodiment, R ²⁹ and R ³¹ are both hydrogen and R ³⁰ is C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl,

And which _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - can be optionally substituted with one or more substituents selected from alkyl,

Wherein R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl,

Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; It can form a 3 to 8 membered heterocyclic ring including.

In a further embodiment, R ²⁹ and R ³¹ are both hydrogen and R ³⁰ is C _4-8 -cycloalkenyl,

And which _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - can be optionally substituted with one or more substituents selected from alkyl,

Where R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl,

Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; It can form a 3 to 8 membered heterocyclic ring including.

In a still further embodiment, R ²⁹ and R ³¹ are both hydrogen and R ³⁰ is cyclohexenyl,

And which _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - can be optionally substituted with one or more substituents selected from alkyl,

Where R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl,

Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; It can form a 3 to 8 membered heterocyclic ring including.

In another embodiment, R ³⁰ is substituted with one C _1-6 -alkyl substituent, such as tert-butyl or methyl.

In yet another embodiment, R ²⁹ , R ³⁰ and R 31 are independently hydrogen, C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl.

In yet another embodiment, R ²⁹ and R ³¹ are both hydrogen and R ³⁰ is C _1-6 -alkyl, C _3-8 -cycloalkyl or C, such as tert-butyl, cyclohexyl or cyclohexenyl _{4-8 -cycloalkenyl} .

In one embodiment, the invention relates to compounds of formula I1:

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , X, D and E are as defined for Formula I or in any of the preceding embodiments. .

In one embodiment, the invention relates to compounds of formula I2:

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , D and E are as defined for Formula I or in any of the preceding embodiments.

In one embodiment, the invention relates to compounds of formula I3:

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ¹⁵ , R ¹⁶ , R ¹⁷ , R ²⁹ , R ³⁰ and R ³¹ are as defined for Formula I, or As defined in any one of the preceding embodiments.

In one embodiment of Formulas I1, I2 and I3, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ and R ⁸ are hydrogen.

In one embodiment, the invention relates to compounds of formula I4:

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , X, D and E are as defined for Formula I or in any of the preceding embodiments. .

In one embodiment, the invention relates to compounds of formula I5:

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁷ , R ⁸ , D and E are as defined for Formula I or in any of the preceding embodiments.

In one embodiment of Formulas I4 and I5, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁷ and R ⁸ are hydrogen.

The compounds of the present invention may have one or more asymmetric centers and are intended to be encompassed within the scope of the present invention by their different optical isomers, when separated, pure or partially purified optical isomers or racemic mixtures.

In addition, geometric isomers can be formed when a double bond or a wholly or partially saturated ring system is present in a molecule. Any geometric isomers, when separated, of pure or partially purified geometric isomers or mixtures are intended to be included within the scope of the present invention. Likewise, molecules containing bonds with limited rotation can form geometric isomers. This is also intended to be included within the scope of this invention.

In addition, some compounds of the present invention may exist in different tautomeric forms and any tautomeric forms that the compounds may form are intended to be included within the scope of the present invention.

The invention also includes pharmaceutically acceptable salts of the compounds. Such salts include pharmaceutically acceptable acid addition salts, pharmaceutically acceptable metal salts, ammonium and alkylated ammonium salts. Acid addition salts include salts of inorganic acids as well as salts of organic acids. Representative examples of suitable inorganic acids include hydrochloric acid, bromic acid, iodic acid, phosphoric acid, sulfuric acid nitric acid, and the like. Representative examples of suitable organic acids include formic acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, propionic acid, benzoic acid, cinnamic acid, citric acid, fumaric acid, glycolic acid, lactic acid, maleic acid, malic acid, malonic acid, mandonic acid, oxalic acid, picric acid, Pyruvic acid, salicylic acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, tartaric acid, ascorbic acid, pamoic acid, bismethylene salicylic acid, ethanedisulfonic acid, gluconic acid, citraconic acid, aspartic acid, stearic acid, palmitic acid, EDTA, glycolic acid, p-aminobenzoic acid, glutamic acid, benzenesulfonic acid, p-toluenesulfonic acid and the like. Further examples of pharmaceutically acceptable inorganic or organic acid addition salts are described in J. Pham. Sci. Pharmaceutically acceptable salts listed in 1977, 66,2. Examples of metal salts include lithium, sodium, potassium, magnesium salts, and the like. Examples of ammonium and alkylated ammonium salts include ammonium, methyl-, dimethyl-, trimethyl-, ethyl-, hydroxyethyl-, diethyl-, n-butyl-, sec-butyl-, tert-butyl-, tetramethylammonium salts and the like. It includes.

Hydrates which the present compounds can form are also intended as pharmaceutically acceptable acid addition salts.

Acid addition salts can be obtained as direct products of compound synthesis. Alternatively, the free base can be dissolved in a suitable solvent containing a suitable acid, and the salt can be separated by evaporating the solvent or otherwise separating the salt from the solvent.

The compounds of the present invention can form solvates with standard low molecular weight solvents using methods well known to those skilled in the art. Such solvates are also intended to be within the scope of the present invention.

The present invention also includes prodrugs of the compounds, which undergo chemical conversion by metabolic processes before they become pharmacologically active substances during administration. In general, such prodrugs can be functional derivatives of compounds of formula (I), which are readily convertible in vivo to the desired compounds of formula (I). Traditional methods for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrug", ed. H. Hundgaard, Elsevier, 1985.

The invention also includes active metabolites of the compounds.

The compounds according to the invention act to antagonize the action of glucagon and are thus useful for the treatment and / or prevention of abnormalities and diseases in which such antagonism is beneficial.

Thus, the present compounds may be used for hyperglycemia, IGT (glucose loading insufficiency), insulin resistance syndrome, syndrome X, type 1 diabetes, type 2 diabetes, hyperlipidemia, dyslipidemia, hypertriglyceridemia, hyperlipoproteinemia, hypercholesterolemia, Applicable to the treatment and / or prevention of atherosclerosis, including gliocytosis, glucagonoma, acute pancreatitis, cardiovascular disease, hypertension, cardiac hypertrophy, gastrointestinal abnormalities, obesity, diabetes due to obesity, diabetic dyslipidemia Can be.

In addition, the present compounds can be applied as a diagnostic agent for identifying patients with defects in glucagon receptor, and as a therapy for increasing gastric acid secretion and dynamics of small intestinal motility due to glucagon administration.

Thus, in a further embodiment the invention relates to a compound according to the invention for use as a drug.

The invention also relates to a pharmaceutical composition comprising at least one compound according to the invention as an active ingredient, together with one or more pharmaceutically acceptable carriers or excipients.

Pharmaceutical compositions are preferred in unit doses comprising about 0.05 mg to about 1000 mg, preferably about 0.1 mg to about 500 mg, and particularly preferably about 0.5 mg to about 200 mg of the compound according to the invention.

Furthermore, the present invention relates to the use of the compounds according to the invention for the preparation of pharmaceutical compositions for the treatment and / or prophylaxis of abnormalities or diseases in which glucagon antagonism is beneficial.

The invention also relates to a method for the treatment and / or prophylaxis of anomalies or diseases in which glucagon antagonism is beneficial, the method comprising administering to a subject in need thereof an effective amount of a compound according to the invention.

In a preferred embodiment of the invention the compound is used in the preparation of a medicament for the treatment and / or prevention of any glucagon-modulated state and disease.

In a preferred embodiment of the invention the compound is used in the preparation of a medicament for the treatment and / or prevention of hyperglycemia.

In a preferred embodiment of the present invention the compound is used for the preparation of a medicament for the treatment and / or prevention of blood sugar reduction in a mammal.

In a preferred embodiment of the invention the compounds are used in the preparation of pharmaceutical compositions for the treatment and / or prophylaxis of IGT.

In a still preferred embodiment of the invention the compounds are used in the preparation of pharmaceutical compositions for the treatment and / or prevention of type 2 diabetes.

In a more preferred embodiment of the invention the compounds are used in the preparation of pharmaceutical compositions for the prolongation and / or prevention of progression from IGT to type 2 diabetes.

In a more preferred embodiment of the invention the compounds are used in the preparation of pharmaceutical compositions for the prolongation and / or prevention of progression from non-insulin demanded type 2 diabetes to insulin demanded type 2 diabetes.

In a further preferred embodiment of the invention the compounds are used in the preparation of pharmaceutical compositions for the treatment and / or prevention of type 1 diabetes. Such treatment and / or prophylaxis is usually combined with insulin therapy.

In a further preferred embodiment of the invention the compounds are used in the preparation of pharmaceutical compositions for the treatment and / or prevention of obesity.

In a further preferred embodiment of the invention the compounds are used in the preparation of pharmaceutical compositions for the treatment and / or prevention of abnormalities in lipid metabolism, such as dyslipidemia.

In a still further preferred embodiment of the invention the compounds are used in the preparation of pharmaceutical compositions for the treatment and / or prevention of appetite control or energy consumption abnormalities.

In a further aspect of the invention the compound is combined with diet and / or exercise.

In an even further aspect of the invention the compound is administered in combination with one or more further active substances in any suitable proportion. Such further active substances may be selected from antidiabetic agents, antihyperlipidemic agents, antiobesity agents, antihypertensive agents and agents for the treatment of complications arising from or associated with diabetes.

Suitable antidiabetic agents are N ^εB29 -tetradecanoyl des (B30) human insulin, EP 214 826 and EP 705 275 (Novo Nordisk A / S) Asp ^B28 human insulin, US disclosed in EP 792 290 (Novo Nordisk A / S), US 5,504,188 (Eli Lilly) Lys ^B28 Pro ^B29 human insulin, insulins, insulin analogs and derivatives such as Lantus of EP 368 187 (Aventis) and GLP-1 derivatives as disclosed in WO 98/08871 (Novo Nordisk A / S) As well as orally active hypoglycemia, all of which are incorporated herein by reference.

Orally active hypoglycemic agents are preferably imidazolines, sulfonylureas, biguanides, meglitinides, oxadiazolidinediones, thiazolidinediones, glucosidase inhibitors, glucagon antagonists, GLP-1 agonists, β-cells Agents acting on ATP-dependent potassium channels of (eg, disclosed in WO 97/26265, WO 99/03861 and WO 00/37474 (Novo Nordisk A / S), incorporated herein by reference, or nateglinide); Potassium channel openers, or potassium channel blockers such as BTS-67582), insulin sensitizers, DPP-IV (dipeptidyl peptidase-IV) inhibitors, PTPase inhibitors, liver enzymes involved in your life and / or glycogenolysis Altering lipid metabolism, lowering food intake, and ALRT-268, such as inhibitors of sugars, sugar absorption activity regulators, GSK-3 (glycogen synthase kinase-3) inhibitors, antihyperlipidemia and antilipidemia With LG-1268 or LG-1069 It is PPAR - and (peroxisome proliferator activated receptor) and RXR include (retinoid X receptor) agonists.

In one embodiment of the invention the compound comprises a N ^εB29 -tetradecanoyl des (B30) human insulin, Asp ^B28 human insulin, Lys ^B28 Pro ^B29 human insulin, Lantus, or a mixed formulation comprising one or more thereof And in combination with insulin or insulin analogues or derivatives.

In a further embodiment of the invention the compound is administered in combination with a sulfonylurea such as tolbutamide, chlorpropamide, tolazamide, glybenclamide, glyburide, glyphide or glycazide do.

In another embodiment of the invention the compound is administered in combination with a biguanide such as metformin.

In yet another embodiment of the invention the compound is administered in combination with a meglitinide such as repaglinide or nateglinide.

In yet another embodiment of the invention the compound is a thiazolidine such as troglitazone, siglitazone, pioglitazone, rosiglitazone, isaglitazone, darglitazone, englitazone, CS-011 / CI-1037 or T174 Dione insulin sensitizers or in combination with compounds disclosed in WO 97/41097, WO 97/41119, WO 97/41120, WO 00/41121 and WO 98/453292 (Dr. Reddy's Research Foundation).

In still another embodiment of the invention the compound is GI 262570, YM-440, MCC-555, JTT-501, AR-H039242, KRP-297, GW-409544, CRE-16336, AR-H049020, LY510929, MBX -102, CLX-0940, insulin sensitizers such as GW-501516 or WO 99/19313, WO 00/50414, WO 00/63191, WO 00/63192, WO 00/63193 (Dr. Reddy's Research Foundation) and WO 00 / 23425, WO 00/23415, WO 00/23445, WO 00/23417, WO 00/23416, WO 00/63153, WO 00/63196, WO 00/63209, WO 00/63190 and WO 00/63189 from Novo Nordisk In combination with a compound disclosed in A / S).

In a further embodiment of the invention the compound is administered in combination with an α-glucosidase inhibitor such as bolgliose, emiglitate, miglitol or acarbose.

In another embodiment of the invention the compound is tolbutamide, chlorpropamide, tolazamide, glybenclamide, glyburide, glyphide, glycazide, BTP-67582, repaglinide or natate It is administered in combination with an agent that acts on ATP-dependent potassium channels of β-cells such as glinides.

In yet another embodiment of the present invention the compound is an antilipidemia or antilipidic agent such as cholestyramine, cholespitol, clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin, probucol or dextyroxine It is administered in combination with a blood thickener.

In another embodiment of the invention, the compound is a combination of metpromin and sulfonylurea, such as glybenclamide or glyburide; Sulfonylureas and acarbose; Metformin and meglitinides such as repaglinide; Acarbose and metformin; Sulfonylureas, metformin and troglitazone; Sulfonylureas, metformin and pioclitazone; Sulfonylureas, metformin and insulin sensitizers such as those disclosed in WO 00/63189 or WO 97/41097; Meglitinides such as repaglinide, metformin and troglitazone; Meglitinides such as repaglinide, metformin and pioglitazone; Meglitinide, metformin, such as repaglinide, and insulin sensitizers, such as those disclosed in WO 00/63189 or WO 97/41097; Insulin and sulfonylureas; Meglitinides such as insulin and repaglinide; Insulin and metformin; Meglitinides such as insulin, metformin and repaglinide; Insulin, metformin and sulfonylurea; Insulin and troglitazone; Insulin and pioglitazone; Insulin and insulin sensitizers such as those disclosed in WO 00/63189 or WO 97/41097; Insulin and lovastatin; Insulin analogues or derivatives, meglitinides such as metformin and repaglinide; Insulin analogues or derivatives, metformin and sulfonylurea; Insulin analogues or derivatives and troglitazone; Insulin analogues or derivatives and pioglitazone; Insulin analogues or derivatives and insulin sensitizers such as those disclosed in WO 00/63189 or WO 97/41097; Insulin analogues or derivatives and lovastatin; And the like in combination with one or more of the compounds mentioned above.

In addition, the compounds according to the invention can be administered in combination with one or more anti-obesity agents or appetite modulators.

Such agents include CART (cocaine amphetamine regulatory transcript) agonists, NPY (neural peptide Y) antagonists, MC4 (melanocortin 4) agonists, orexin antagonists, TNF (tumor necrosis factor) activity modulators, CRF (corticotropin secretion factor Agonists, CRF BP (corticotropin secretion factor binding protein) antagonists, urocortin agonists, CL-316243, AJ-9677, GW-0604, LY362884, LY377267 or AZ-40140 agonists, MSH (melanocytes) Stimulating hormone) agonists, MCH (melanocyte-aggregating hormone) antagonists, CCK (cholecystokinin) agonists, serotonin reuptake inhibitors such as fluoxetine, seroxat or citalopram, serotonin and noradrenaline reuptake inhibitors, 5HT (serotonin) agonists , Bombesin agonists, galanin antagonists, growth hormones, growth hormone secreting compounds, TRH (thyretropin secreting hormone) agonists, UCP 2 or 3 (uncoupling protein 2 or 3) activity modulators, rep Tin agonists, DA (dopamine) agonists (bromocriptine, doprexin), lipase / amylase inhibitors, PPAR modulators, RXR modulators or TR β agonists.

In one embodiment of the invention the anti-obesity agent is leptin.

In another embodiment of the invention the anti-obesity agent is dexampetamine or amphetamine.

In another embodiment of the invention the anti-obesity agent is fenfluramine or dexfenfluramine.

In yet another embodiment of the invention the antiobesity agent is sibutramine.

In a further embodiment of the invention the anti-obesity agent is orlistat.

In another embodiment of the invention the anti-obesity agent is malzindol or phentermine.

In addition, the compounds may be administered in combination with one or more antihypertensive agents. Examples of antihypertensive agents include β-blockers such as alprenolol, atenolol, tinolol, pindolol, propranolol and metoprolol, benazepril, captopril, enalapril, posinopril, ricinopril, quinapril and ramipril and Calcium channel blockers, such as ACE (angiotensin converting enzyme) inhibitors, nifedipine, felodipine, nicardipine, isradipine, nimodipine, diltiazem and verapamil, and doxazosin, urapidil, prazosin and terrazocin α-blockers. Further reference is Remington: The Science and Practice of Pharmacy, 19th Edition, Gennaro, Ed., Mack Publishing Co., Easton, PA, 1995.

It is to be understood that any suitable combination of a compound according to the invention with a diet and / or exercise, one or more of the above mentioned compounds and optionally one or more other active substances is contemplated within the scope of the invention.

Pharmacological composition

The compounds of the present invention may be administered in single or multiple doses, alone or in combination with pharmaceutically acceptable carriers or additives. Pharmaceutical compositions according to the present invention are disclosed in Remington: The Science and Practice of Pharmacy, 19th Edition, Gennaro, Ed., Mack Publishing Co., Easton, PA, 1995, as well as pharmaceutically acceptable carriers or diluents. It may be formulated with any other known auxiliaries and additives according to traditional techniques such as.

The pharmaceutical compositions may be oral, rectal, nasal, lung, topical (including buccal and sublingual), transdermal, intravaginal, intraperitoneal, vaginal and subcutaneous, intramuscular, intramedullary, intravascular and intradermal) Specifically formulated for administration by any suitable route, such as parenteral route, oral route is preferred. It should be appreciated that the preferred route depends on the general condition and age of the subject being treated, the condition being treated and the characteristics of the active ingredient selected.

Pharmaceutical compositions for oral administration include solid dosage forms such as capsules, tablets, dragees, pills, lozenges, powders, and granules. To serve any purpose, it may be formulated with a coating, such as an enteric coating, or formulated to provide controlled release of the active ingredient, such as sustained or sustained release according to methods well known in the art.

Liquid dosage forms for oral administration include solutions, emulsions, suspensions, syrups and elixirs.

Pharmaceutical compositions for parenteral administration include sterile aqueous and non-aqueous injectable solutions, sprays, suspensions or emulsions, as well as sterile powders that are re-liquefied into sterile injectable solutions or sprays prior to use. Depot injectable formulations are also intended to be within the scope of this invention.

Other suitable dosage forms include suppositories, sprays, ointments, creams, gels, inhalants, dermal adherents, transplants, and the like.

Typical oral dosages range from about 0.001 to about 100 mg / kg per day, preferably from about 0.01 to about 50 mg / kg per day, more preferably from about 0.05 to about 10 mg / kg per day, eg For example, in dosages of 1-3. The exact dosage depends on the frequency and mode of administration, the sex, age, weight and general condition of the subject being treated, the condition being treated and the nature and severity of any accompanying disease to be treated, and other factors demonstrated to those skilled in the art.

The formulations may conveniently be presented in unit dosage form by methods well known to those skilled in the art. Typical unit dosage forms for oral administration once or more, for example 1-3 times, may contain from 0.05 to about 1000 mg, preferably from about 0.1 to about 500 mg, more preferably from about 0.5 mg to about 200 mg. .

For parenteral routes such as intravascular, intramedullary, intramuscular and similar administration, typical dosages are about half of those employed for oral administration.

The compounds of the present invention are generally used as free substances or pharmaceutically acceptable salts thereof. One example is base addition salts of compounds with the use of free acids. When the compound of formula (I) contains free acid, such salts are prepared by conventional methods by treating a solution or suspension of free acid (I) with a chemical equivalent of a pharmaceutically acceptable base. Representative examples are mentioned above.

For parenteral administration, solutions of novel compounds of formula (I) in sterile aqueous solutions, aqueous propylene glycol, aqueous vitamin E, sesame oil or peanut oil can be employed. This aqueous solution should be suitably buffered if necessary and the liquid diluent firstly isotonic with sufficient saline or glucose. Aqueous solutions are particularly suitable for intravascular, intramuscular, subcutaneous and intraperitoneal administration. Adopted sterile aqueous media is readily available by standard techniques known to those skilled in the art.

Suitable pharmaceutical carriers include inert solid diluents or fillers, sterile aqueous solutions and various organic solvents. Examples of solid carriers are lower alkyl zateres of lactose, clay, sucrose, cyclodextrin, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and cellulose. Examples of liquid carriers are syrup, peanut oil, olive oil, phospholipids, fatty acids, fatty acid amines, polyoxyethylene and water. Similarly, the carrier or diluent may comprise any oleaginous secretory material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or in admixture with wax. Thereafter, the pharmaceutical composition formed by the combination of the novel compound of formula I and a pharmaceutically acceptable carrier is easily administered in various dosage forms suitable for the disclosed route of administration. The formulations may conventionally appear in unit dosage form by methods known in the art of pharmacy.

Formulations of the invention suitable for oral administration may be presented in discrete units such as capsules or tablets, each containing a predetermined amount of the active ingredient, which may include suitable additives. In addition, orally available formulations may be in the form of powders or granules, solutions or suspensions in aqueous or non-aqueous liquids, or oil-in-water or water-in-oil liquid emulsions.

If a solid carrier is used for oral administration, it can be formulated or placed in hard gelatin capsules in powder or pellet form, or in the form of troches or lozenges. The amount of solid carrier can vary widely but usually ranges from about 25 mg to about 1 g. If a liquid carrier is used, it may be formulated in the form of a sterile injectable liquid, such as a syrup, emulsion, light gelatin capsule or aqueous or non-aqueous liquid suspension or solution.

Typical tablets that may be formulated by traditional tableting techniques may include the following.

main point:

Active compound (free compound or salt thereof) 5.0 mg

Lactosum Ph. Eur. 67.8 mg

Cellulose, microcrystalline. (Avicel) 31.4 mg

Amberlite® IRP88 * 1.0 mg

Magnesii stearas Ph. Eur. q. s.

coating:

About 9 mg of hydroxypropyl methylcellulose

Mywacett 9-40 T ** approx.0.9 mg

Polacrillin potassium NF, tablet disintegrant, Rohm and Haas.

** Acylated monoglycerides used as plasticizers for film coating.

If desired, the pharmaceutical compositions of the present invention may comprise a compound of formula (I) in combination with further pharmaceutically active substances as described above.

The following examples and general methods refer to intermediate compounds and end products identified in the specification and the synthetic schematics. Although the preparation of the compounds of the present invention is described in detail using the following examples, the chemical reactions described are disclosed in terms of their general applicability to the preparation of the glucagon antagonists of the present invention. At times, the reaction may not be applicable as described for each compound included within the disclosed scope of the invention. Compounds so produced can be readily recognized by those skilled in the art. In this case the reaction can be carried out successfully with traditional modifications known to those skilled in the art, ie, proper protection of the intervening groups, changes to other traditional reagents, or customary modifications of the reaction state. Alternatively, other reactions disclosed or otherwise conventional may be applicable to the preparation of the corresponding compounds of the present invention. In all preparation methods, all starting materials are known or can be readily prepared from known starting materials. All temperatures are in degrees Celsius and otherwise indicated, all percentages and percentages when referring to yield are by mass and all parts when referring to solvents and eluents are by volume.

Some NMR data shown in the following example are just selected data.

In the examples and pharmaceutical methods, the following terms are intended to have the following meanings:

DCM: Dichloromethane

DMF: N, N-dimethylformamide

DMSO: Dimethyl Sulfoxide

M.p .: Melting Point

TFA: trifluoroacetic acid

THF: tetrahydrofuran

EDAC: 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochlor

Ride

HOBt: 1-hydroxybenzotriazole

HOAt: 3-hydroxy-3H- [1,2,3] triazolo [4,5-b] pyridine

Also referred to as 1-hydroxy-7-azabenzotriazol

EGTA: ethylene glycol bis (β-aminoethyl ether) N, N, N'N'-tria

Set mountain

BSA: N, O-bis (trimethylsilyl) acetimidadate

IBMX: isobutylmethylxanthine

HPLC-MS (Method A)

The following instruments were used:

ㆍ Sciex API 100 Single Quadrupole Mass Spectrometer

ㆍ Perkin Elmer Series 200 Quard Pump

ㆍ Perkin Elmer Series 200 Automatic Sampler

ㆍ Applied Biosystems 785A UV Detector

Sedex 55 Evaporative Light Scattering Detector

Valco column switch with Valco actuator controlled by a timed event from the pump

The Sciex Sample control software was run on a Macintosh PowerPC 7200 computer for machine control and data acquisition.

The HPLC pump was connected to four eluent reservoirs containing:

A: acetonitrile

B: water

C: 0.5% TFA in water

D: 0.02M ammonium acetate

The requirement for the sample is that it contains about 500 μg / ml of the compound for analysis in an acceptable solvent such as methanol, ethanol, acetonitrile, THF, water and a mixture of compounds. May interfere with chromatography at low acetonitrile concentrations.)

The assay was performed at room temperature by injecting 20 μl of sample solution into the column and eluted with a gradient of acetonitrile in 0.05% TFA or 0.002 M ammonium acetate. Various elution conditions were used depending on the analytical method.

The eluate from the column passed through a flow splitting T-connector, which passed through an API interface of API 100 spectrometer through an approximately 1 m.75 µ fused silica capillary at a rate of approximately 20 μL / min. It was.

The remaining 1.48 mL passed through the UV detector to the ELS detector.

Detection data were obtained simultaneously from the mass spectrometer, UV-detector and ELS detector during LC-analysis.

The LC states, detector settings and mass spectrometer settings used in the different methods are given in the following table.

column YMC ODS-A120Ås-5μ 3mm X 50mm id gradient At 1.5mL / min Linearly 5% to 90% acetonitrile in 0.05% TFA for 7.5 min detection UV: 214 nm ELS: 40 ℃ MS Experiment: Start: 100amu Stop: 800 amu Step: 0.2 amu Pause: 0.571msec Method: Scan 284 times = 9.5 min

HPLC-MS (Method B)

The following instruments were used:

ㆍ Hewlett Packard series 1100 G1312A Bin Pump

ㆍ Hewlett Packard series 1100 Column section

Hewlett Packard series 1100 G13 15A DAD Diode Alignment Detector

Hewlett Packard series 1100 MSD

The machine was controlled by HP Chemstation software.

The HPLC pump was connected to two eluent reservoirs containing:

A: 0.01% TFA in water

B: 0.01% TFA in acetonitrile

The assay was performed at 40 ° C. by injecting an appropriate volume of sample (preferably 1 μl) onto the column and eluted with a gradient of acetonitrile.

The LC conditions, detector settings and mass spectrometer settings used are given in the following table.

column Waters XterraMS C-18 X 3 mm id gradient Straight at 1.0 mL / min 10% to 100% acetonitrile for 7.5.min detection UV: 210 nm (analog output from the ADC) MS Ionization Method: API-ESScan 100 to 1000 amu steps 0.1 amu

HPLC-MS (Method C)

The following instruments were used:

ㆍ Hewlett Packard series 1100 G1312A Bin Pump

Hewlett Packard series 1100 G13 15A DAD Diode Alignment Detector

ㆍ Sciex 300 Triple Quadrupole Mass Spectrometer

ㆍ Gilson 215 Microinjector

Sedex 55 Evaporative Light Scattering Detector

The MassChrom 1.1.1 software was run on a Macintosh G3 computer to control the pump and detector. Gilson Unipoint Version 1.90 controls the autoinjector.

The HPLC pump was connected to two eluent reservoirs containing:

A: 0.01% TFA in water

B: 0.01% TFA in acetonitrile

The assay was performed at room temperature by injecting an appropriate volume of sample (preferably 1 μl) onto the column, eluting with a gradient of acetonitrile.

The LC conditions, detector settings and mass spectrometer settings used are given in the following table.

column YMC ODS-A120Ås-5μ 3mm X 50mm id gradient Straight at 1.5 mL / min 5% to 90% acetonitrile for 7.5.min detection 210 nm (analog from DAD Print) MS Ionization Method: API-ESScan 100 to 1000 amu steps 0.1 amu

Preparation of building blocks used in the examples below

Building block 1: (RS) -isoserine ethyl ester hydrochloride

Anhydrous ethanol (40 mL) was cooled in an ice bath and thionyl chloride (4 mL) was added dropwise while maintaining the temperature below 5 ° C. (RS) -isoserine (2.5 g, 23.79 mmol) was added to this cooling solution, and stirring was continued until a homogeneous solution was obtained.

The ice bath was removed and stirring continued at room temperature for 17 hours. The solution was concentrated in vacuo to yield 4.0 g (100%) of (RS) -isoserine ethyl ester hydrochloride in oil form.

¹ H-NMR (DMSO- d ₆ ): δ1.22 (t, 3H), 3.00 (dm, 2H), 4.15 (q, 2H), 4.40 (dd, 1H), 6.30 (br s, 1H), 8.32 (br s, 2 H). _{^{13 C-NMR (DMSO- d 6}} ): δ14.7 (q), 42.4 (t), 61.7 (t), 67.7 (d), 171 (s).

Building block 2: (R) -isoserine ethyl ester hydrochloride

Step A: (R)-(2,2-Dimethyl-5-oxo- [1,3] dioxolan-4-yl) acetic acid

To a suspension of D-(+)-malic acid (15.0 g, 0.1119 mol) in toluene anhydrous (150 mL) was added 2,2-dimethoxypropane (50 mL, 0.392 mmol). The mixture was refluxed at 100 ° C. for 2 hours and then evaporated in vacuo. The residue was dissolved in diethyl ether (150 mL) and subjected to flash column chromatography using diethyl ether (200 mL) as eluent. The pure portions were combined and evaporated in vacuo and the residue was stirred in n-hexane. The precipitate was collected, washed with n-hexane and dried to give 15.7 g (81%) of (R)-(2,2-dimethyl-5-oxo- [1,3] dioxolan-4-yl) acetic acid as a solid. Obtained in form.

¹ H-NMR (acetone- d ₆ ): δ 1.57 (ds, 6H), 2.85 (m, 2H), 4.80 (dd, 1H), 11.0 (br s, 1H). ¹³ C-NMR (acetone- d ₆ ): 8 25.9 (q), 26.8 (q), 36.2 (t), 71.5 (d), 111.2 (s), 170.7 (s), 172.7 (s).

Microanalytical Theoretical Values for C ₇ H ₁₀ C ₅ :

C, 48.28%; H, 5.79%. Found:

C, 48.31%; H, 6.09%.

Step B: (R)-(2,2-Dimethyl-5-oxo- [1,3] dioxolan-4-ylmethyl) carbamic acid benzyl ester

(R)-(2,2-dimethyl-5-oxo- [1,3] dioxalan-4-yl) acetic acid (10.0 g, 57.41 mmol) in anhydrous toluene (100 mL), triethylamine (10 mL, 68.89 mmol) and diphenylphosphoryl azide (14 mL, 63.15 mmol) were heated and stirred at 85 ° C. Stirring was continued for an additional 1 hour when gas evolution started. Anhydrous benzyl alcohol (6.3 mL, 63.15 mmol) was added and heating continued for 17 h. After evaporation in vacuo, the residue was partitioned into dichloromethane, water and brine. The aqueous phase was extracted twice more with dichloromethane. The combined organic phases were washed twice with saturated sodium hydrogen carbonate. After drying (magnesium sulfate), filtration, and concentration of the organic phase in vacuo, the residue was flash column chromatography using dichloromethane as eluent. 6.4 g (40%) of (R)-(2,2-dimethyl-5-oxo- [1,3] dioxalan-4-ylmethyl) carbamic acid benzyl ester was obtained in the form of an oil.

¹ H-NMR (acetone- d ₆ ): δ1.56 (s, 6H), 3.61 (m, 2H), 4.64 (dd, 1H), 5.08 (dd, 2H), 6.51 (br s, 1H), 7.29 -7.38 (m, 5 H).

Step C: (R) -3-benzyloxycarbonylamide-2-hydroxypropionic acid

(R)-(2,2-dimethyl-5-oxo- [1,3] dioxolan-4-ylmethyl) carbamic acid benzyl ester (6.0 g, 25.08 mmol) in acetonitrile (100 mL) was dissolved in hydrochloric acid (1N, 100 mL). The mixture was stirred at 40 ° C. for 3 hours and concentrated to half the original volume. The solid was collected by filtration and washed with water. The crude product in acetone (100 mL) was stirred for 5 minutes and filtered. Toluene was added to the colorless clear filtrate and concentrated in vacuo until a precipitate was obtained. The precipitate was collected by filtration and dried to afford 4.45 g (87%) of (R) -3-benzyloxycarbonylamino-2-hydroxypropionic acid.

¹ H-NMR (acetone- d ₆ ): δ 3.45 (ddd, 1 H), 3.58 (ddd, 1 H), 4.29 (dd, 1 H), 5.08 (s, 2H), 6.41 (br s, 1H ), 7.29-7.38 (m, 5 H). ¹³ C-NMR (acetone- d ₆ ): δ 45.1 (t), 66.4 (t), 70.4 (d), 128.3 (d), 128.9 (d), 138.0 (d), 157.2 (s), 173.8 ( s); HPLC-MS (Method C): m / z = 262 (M + Na ⁺ ); R _t = 2.20 min. M. p. 131-132 ° C.

Microanalytical Theoretical Values for C ₁₁ H ₁₃ NO ₅ :

C, 55.23%; H, 5.48%; N, 5.85%. Found:

C, 55.35%; H, 5.72%; N, 5.82%.

Step D: (R) -Isoserine

(R) -3-benzyloxycarbonylamino-2-hydropropionic acid (4.4 g, 18.39 mmol) was dissolved in anhydrous ethanol (150 mL). Palladium (10%, 0.5 g) in activated carbon under nitrogen atmosphere was added and the mixture was hydrogenated at 1 atmosphere for 17 hours. The catalyst was removed by filtration and washed with water. The combined filtrate and washes were concentrated to about 20 mL by vacuum evaporation. A precipitate was obtained by the dropwise addition of methanol (100 mL). The precipitate was filtered off, washed with methanol and dried to yield 1.78 g (92%) of (R) -isoserine in solid form.

¹ H-NMR (D ₂ 0): δ 3.07 (dd, 1H), 3.30 (dd, 1H), 4.19 (dd, 1H). ¹³ C-NMR (D ₂ 0): δ 43.0 (t), 68.9 (d), 177.5 (s). Mp 200-201 ° C.

Step E: (R) -isoserine ethyl ester hydrochloride

Compounds were prepared analogously to the method outlined above for (RS) -isoserine ethyl ester hydrochloride.

¹ H-NMR (DMSO- d ₆ ): δ1.22 (t, 3H), 2.88 (dd, 1H), 3.10 (dd, 1H), 4.14 (q, 2H), 4.40 (m, 1H), 6.32 ( d, 1 H), 8.28 (br s, 2 H). _{^{13 C-NMR (DMSO- d 6}} ): δ13.9 (q), 41.4 (t), 60.7 (t), 66.9 (d), 170.9 (s).

Building Block 3: (S)-(2,2-Dimethyl-5-oxo- [1,3] dioxolan-4-ylmethyl) -4-ylmethylammonium trifluoroacetate

Step A: (S)-(2,2-Dimethyl-5-oxo- [1,3] dioxolan-4-yl) acetic acid

To a suspension of L-malic acid (3 g, 22.4 mmol) in toluene (25 mL) was added 2,2-dimethoxypropane (8.5 g, 81 mmol). The suspension was heated to reflux for 20 minutes. The solvent was evaporated in vacuo to afford (S)-(2,2-dimethyl-5-oxo [l, 3] -4-yl) -acetic acid.

_{^{1 H-NMR (DMSO- d 6}} ): δ1.52 (6H, d), 2.75 (2H, t), 4.78 (1H, t), 12.52 (1H, br s).

Step B: (S)-(2,2-Dimethyl-5-oxo- [1,3] dioxolan-4-ylmethyl) carbamic acid benzyl ester

(S)-(2,2-dimethyl-5-oxo- [1,3] dioxolan-4-yl) acetic acid (1 g, 5.7 mmol) and triethylamine (0.66 g, 6.5 mmol) in toluene (20 mL) Was added to phosphorazide acid diphenyl ester (1.7 g, 6.2 mmol). The solution was heated to reflux for 1 hour. Benzyl alcohol (0.54 g, 5 mmol) was added and maintained at reflux for an additional 6 hours. After cooling, the solution was partitioned between ethyl acetate (2 x 50 mL) and aqueous sodium hydrogen carbonate solution (10%, 2 x 50 mL). The organic layer was collected, dried (sodium sulfate), and the solvent was removed by vacuum evaporation to give 976 mg of crude (S)-(2,2-dimethyl-5-oxo- [1,3] dioxolan-4-ylmethyl) carbox Chestnut acid benzyl ester was obtained in oil form.

¹ H-NMR (DMSO- d ₆ ): δ 1.53 (6H, d), 3.34 (1H, m), 3.50 (1H, m), 4.50 (1H, d), 4.64 (1H, t), 5.03 ( 2H, d), 7.32 (5H, m)

Step C: (S) -2,2-Dimethyl-5-oxo- [1,3] dioxolan-4-ylmethylammonium trifluoroacetate

Crude (S)-(2,2-dimethyl-5-oxo- [1,3] dioxalan-4-ylmethyl) carbamic acid benzyl ester (976 mg, 3.5 mmol) was dissolved in ethanol (14 mL) and then palladium (10% on activated charcoal, 300 mg) and 1,3-cyclohexadiene (2.8 g, 35 mmol) were added to stir the reaction at room temperature for 1 hour and heated to 40 ° C. for 10 minutes. After filtration, TFA (0.4 g, 3.5 mmol) was added and the solvent was evaporated to remove and then crude (S) -2,2-dimethyl-5-oxo- [1,3] dioxalan-4-ylmethylammonium trifluor Low acetate was obtained in the form of an oil.

¹ H-NMR (CDCl ₃ ): δ 1.46 (6H, d), 3.40-3.70 (2H, m), 4.38 (1H, m), 7.25 (> 4H, br s); HPLC-MS (Method B): m / z = 146 (M ⁺ ); R _t = 0.38 min.

Building Block 4: 3-Amino-2-fluoropropionic Acid Methyl Ester

In an ice bath anhydrous methanol (5.3 mL) was cooled to -15 ° C and thionyl chloride (2.5 mL) was added dropwise while maintaining the temperature below 5 ° C. (RS) -3-Amino-2-fluoropropionic acid (0.27 g, 2.52 mmol) was added to the cooled solution and stirring continued until a homogeneous solution was obtained. The ice bath was removed and stirring continued for 17 hours at room temperature. The solution was concentrated in vacuo and co-evaporated three more times with anhydrous methanol. The residue was filtered off, washed with DCM and dried to give 0.13 g (33%) of 3-amino-2-fluoropropionic acid methyl ester hydrochloride in solid form.

¹ H-NMR (DMSO- ₆ d): δ3.36 (m, 2H), 3.76 (s, 3H), 5.51 (d, 2H), 8.59 (br s, 3H).

Building Block 5: 3-Amino-2 (R) -methoxypropionic acid methyl ester hydrochloride

Step (A): (R) -2-hydroxysuccinic acid dimethyl ester

Acetyl chloride (12.5 mL) was added to an ice cooled methanol solution (250 mL), and the solution was stirred at 0 ° C. for 1 h. (R) -malic acid (20.0 g) was added and the solution stirred for 16 h at room temperature. The solvent was evaporated in vacuo to give quantitative yield of (R) -2-hydroxysuccinic acid dimethyl ester in oil form.

¹ H-NMR (CDCl ₃ ): delta 4.52 (dd, 1H), 3.80 (s, 3H), 3.71 (s, 3H), 3.55 (bs, 1H), 2.88 (dd, 1H), 2.80 (dd, 1H).

Step (B): (R) -2-methoxysuccinic acid dimethyl ester

The above (R) -2-hydroxysuccinic acid dimethyl ester was redissolved in methyl iodine (100 mL), and freshly prepared silver oxide (30.2 g) was added and the mixture was stirred at room temperature for 24 hours. The reaction mixture was diluted with acetonitrile (200 mL) and filtered through celite to remove salts of silver and excess silver oxide. The filtrate was dried to give (R) -2-methoxysuccinic acid dimethyl ester (23.2 g, 88%) in oil form.

¹ H-NMR (CDCl ₃ ): δ 4.20 (dd, 1H), 3.78 (s, 3H), 3.71 (s, 3H), 3.48 (s, 3H), 2.80 (dd, 2H).

Step (C): (R) -2-methoxysuccinic acid 1-methyl ester

The above (R) -2-methoxysuccinic acid dimethyl ester was suspended in 2N aqueous hydrochloric acid, heated to reflux for 30 minutes to give a clear solution. After evaporation of the solvent in vacuo 2 (R) -methoxysuccinic acid was obtained in the form of an oil. The oil was redissolved in acet anhydride (120 mL) and heated to 110 ° C. for 2 hours. The solvent was removed by rotary evaporation to give an oil. Ice cooled methanol (150 mL) was added and the mixture was stirred at 0 ° C. for 3 hours and then at room temperature for 16 hours. The solvent was removed to obtain (R) -2-methoxysuccinic acid 1-methyl ester.

¹ H-NMR (CDCl ₃ ): δ 10.30 (bs, 1H), 4.19 (dd, 1H), 3.80 (s, 3H), 3.50 (s, 3H), 2.86 (dd, 1H), 2.78 (dd, 1 H).

Step (D): 3- tert Butoxycarbonylamino-2 (R) -methoxypropionic acid methyl ester

Without further purification, the above (R) -2-methoxysuccinic acid 1-methyl ester (5.0 g, 30.8 mmol) was dissolved in thionyl chloride (16 mL) and heated to reflux for 2 h. Thionyl chloride and its traces were removed by rotary evaporation followed by co-evaporation with acetonitrile.

^OneH-NMR (CDCl₃): delta 3.27 (dd, 1H), 3.48 (dd, 1H), 3.51 (s, 3H), 3.80 (s, 3H), 4.22 (dd, 1H).

Pure acid chloride was dissolved in toluene (50 mL). Trimethylsilylazide (5.0 mL, 38.2 mmol) was added and the mixture was heated at 100 ° C. overnight. Then tert-butanol (30 mL) was added and heating continued for an additional 16 hours. The reaction mixture was cooled down and undissolved material was removed by filtration. The organic phase was washed with water (100 mL), saturated sodium bicarbonate solution (100 mL), 10% citric acid solution (100 mL), water (100 mL) and saturated sodium chloride solution (100 mL) and then dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residual oil was further purified by column chromatography using 20% ethyl acetate / heptane as eluent. Pure portions (TLC plates were dyed with ammonium molybdate / cerium sulfate / sulfonic acid) were dried. The final yield of 3-tert-butoxy-carbonylamino-2 (R) -methoxypropionic acid methyl ester was 600 mg (9%).

¹ H-NMR (CDCl ₃ ): δ 6.63 (t, 1H), 3.83 (t, 1H), 3.64 (s, 3H), 3.25 (s, 3H), 3.18 (dd, 2H), 1.36 (s, 9H).

Step (E): 3-Amino-2 (R) -methoxypropionic acid methyl ester hydrochloride

3-tert-butoxycarbonylamino-2-methoxypropionic acid methyl ester (500 mg, 2 mmol) was dissolved in 10% TFA in DCM (20 mL) and the reaction mixture was stirred at ambient temperature for 30 minutes. The solvent was removed and the residue was co-evaporated twice from 30 mL of IN hydrochloric acid in ether. Yield: 320 mg (88%).

¹ H-NMR (CDCl ₃ ): δ 8.25 (s, 3H), 4.21 (dd, 1H), 3.71 (s, 3H), 3.40 (s, 3H), 3.15 (m, 1H), 2.98 (m, 1H).

Building Block 6: (R) -3- (9H-fluorene-9-ylmethoxycarbonylamino) -2-hydroxypropionic acid ((R) -Fmoc-isoserine

Toluene (100 mL), triethylamine (4.7 mL, 33.8 mmol) in (R)-(2,2-dimethyl-5-oxo [1,3] dioxalan-4-yl) acetic acid (5.88 g, 33.8 mmol) ) And diphenoxyphosphoryl azide (8.0 mL, 37.2 mmol) were added. The reaction mixture was heated to 100 ° C. and stirred at rt for 1.5 h. 9-fluorenemethanol (5.1 g, 26 mmol) was added and the reaction mixture was refluxed for 6 hours. After cooling to room temperature the mixture was transferred to a separatory funnel and washed with water (2x50 mL). The solvent was removed in vacuo and co-evaporated with acetonitrile (100 mL). The remaining light brown oil was dissolved in DCM (20 mL) and purified on silica gel column using DCM as eluent. DCM was evaporated off to give a pale yellow oil which was redissolved in acetonitrile (150 mL) and aqueous hydrochloric acid (1N, 75 mL) was added. The yellow reaction mixture was stirred at rt for 3 h. The solvent was removed in vacuo, toluene (100 mL) was added and the suspension was heated to reflux and allowed to cool to room temperature. (R) -Fmoc-isoserine (3.1 g, 28%) was isolated in powder form by filtration. Mp: 165-166 ° C ..

¹ H-NMR (DMSO- d ₆ ): δ 12.5 (br s, 1 H), 7.90 (m, 2 H), 7.70 (m, 2 H), 7.50-7.30 (m, 4 H), 5.45 (br s , 1H), 4.23 (m, 3H), 4.05 (m, 1H), 3.17 (m, 1H); HPLC-MS (Method B): m / z = 350 (M + Na); R _t = 3.20 min.

Building Block 7: 3- (tert-Butyldimethylsilanyloxymethyl) -4-trifluoromethoxyphenyl amine

Step A: 5-nitro-2-trifluoromethoxybenzoic acid methyl ester

In a three-necked round bottom flask equipped with thermometer and separatory funnel, HNO ₃ (5 mL fumes, 100%) was cooled in an ice bath. Methyl 2- (trifluoromethoxy) benzoate (5 g, 22.7 mmol) was added slowly to the cooled HN03 within 0.5 hours while maintaining the temperature below 15 ° C. The reaction was then stirred at 60 ° C. for 1 hour and at room temperature for 2 hours. The mixture was added to ice to separate the oil. Water (50 mL) was added to the oily residue, neutralized with aqueous sodium hydrogen carbonate solution, and extracted with ethyl acetate (25 mL). The aqueous phase was extracted one more time with ethyl acetate (15 mL). The combined organic phases were washed with saturated sodium chloride (2 x 15 mL), dried (magnesium sulfate) and concentrated in vacuo to give 5.69 g of 5-nitro-2-trifluoromethoxybenzoic acid methyl ester.

_{^{1 H-NMR (DMSO- d 6}} ): δ3.93 (3H, s), 7.82 (1H, d), 8.58 (1H, d), 8.67 (1H, s).

Step B: 5-Amino-2-trifluoromethoxybenzoic acid methyl ester

5-nitro-2-trifluoromethoxybenzoic acid methyl ester (5.69 g, 21.5 mmol) was dissolved in 99.9% (80 mL) of ethanol and tin (II) chloride dihydrate (24.2 g, 107 mmol) was added. The suspension was stirred at 75 ° C. for 2 hours and then concentrated in vacuo. To the residue was added ethyl acetate (100 mL) and water (50 mL) and adjusted to pH 8 with 4N sodium hydroxide (50 mL). The liquid was removed from the resulting fine precipitate and then washed twice with ethyl acetate. The combined organic phases were washed with water: saturated sodium chloride (1: 1) solution (2 × 100 mL), dried (magnesium sulfate) and concentrated in vacuo. The residue was purified by column chromatography (120 g silica) using ethyl acetate: heptane (1: 1) as eluent to afford 3.8 g of 5-amino-2-trifluoromethoxybenzoic acid methyl ester.

_{^{1 H-NMR (DMSO- d 6}} ): δ3.82 (3H, s), 5.63 (2H, s), 6.79 (1H, d), 7.07 (1H, s), 7.11 (1H, d).

Step C: (5-amino-2-trifluoromethoxyphenyl) methanol

5-amino-2-trifluoromethoxybenzoic acid methyl ester (3.0 g, 12.8 mmol) was dissolved in THF (20 mL) in a three-necked flask equipped with a thermometer and a separatory funnel under a nitrogen atmosphere. In combination with stirring and ice-cooling, lithium aluminum hydride (1M in THF, 15 mL) was added dropwise over 10 minutes. Stirring was continued for 1 hour at room temperature, and the reaction mixture was concentrated in vacuo. The residue was suspended in DCM (150 mL) and water (50 mL) and filtered through celite. The filtrate was partitioned between DCM and water. The combined organic phases were washed with water (2x20 mL), dried (magnesium sulfate) and concentrated in vacuo to yield 2.47 g of (5-amino-2-trifluoromethoxyphenyl) methanol.

_{^{1 H-NMR (DMSO- d 6}} ): δ3.92 (2H, d), 5.18 (1H, t), 5.28 (2H, s), 6.45 (1H, d), 6.91 (1H, d).

Step (D): 3- (tert-butyldimethylsilanyloxymethyl) -4-trifluoromethoxyphenylaniline

(5-amino-2-trifluoromethoxyphenyl) methanol (1.2 g, 5.8 mmol) was dissolved in DMF (5 mL), imidazole (0.48 g, 7.1 mmol) and tert-butyldimethylsilyl chloride (0.99 g, 6.6 mmol) was added and the mixture was stirred for 16 h. The reaction mixture was extracted with ethyl acetate (50 mL) and water (20 mL). The aqueous phase was extracted once more with ethyl acetate (20 mL). The combined organic phases were washed with water (10 mL), aqueous citric acid (10 mL, 10%) and water (2 × 10 mL), dried (magnesium sulfate) and concentrated in vacuo. Purify the residue by column chromatography (110 g, silica gel) using ethyl acetate and heptane (1: 3) as eluent to obtain 1.2 g of 3- (tert-butyldimethylsilanyloxymethyl) -4-trifluoromethoxy. Phenylaniline was obtained.

^OneH-NMR (DMSO-d ₆ ) : δ 0.82 (9H, s), 3.25 (6H, s), 4. 52 (2H, s), 5.23 (2H, s), 6.41 (1H, d), 6.61 (1H, s), 6.86 (1H, d).

Building Block 8: 4-cyclohex-1-enylaniline

This compound is described in J. v. Similar to those described in Braun et al ., J. Liebigs Ann . Chem ., 472 (1929), 1-89, aniline (2 equivalents) in ethanol and 37% hydrochloric acid, cyclohexanone (1 Equivalent) was prepared from addition of ethyl acetate, water, and sodium hydroxide, neutralization with 85% phosphoric acid, phase separation and distillation of the organic phase. The residue was added to the catalytic amount of sulfuric acid and distilled (180 ° C., 5-7 mbar). The distillate was re-distilled (120 ° C., 3 mbar) (in the residue) to afford the desired 4-cyclohex-1-enylaniline.

¹ H NMR (DMSO- d ₆ ): δ1.50-1.60 (m, 2H), 1.60-1.70 (m, 2H), 2.10-2.15 (m, 2H), 2.20-2.30 (brd s, 2H), 5.00 (s, 2H), 5.90 (t, 1H), 6.50 (d, 2H), 7.10 (d, 2H).

Building Block 9: 4-cyclohexylaniline

This compound is commercially available (for example from Lancaster or Avocado).

Building Block 10: 4-cyclohexylcyclohexylamine

The preparation of this compound is described in H. Booth et al., J. Chem. Soc . (B), 1971, 1047-1050.

Building Blocks 11: 4- (2-methylcyclohex-1-enyl) aniline and (R, S) -4- (6-methylcyclohex-1-enyl) aniline

A mixture of 2-methyl-cyclohexanone (112 g, 1.0 mol), aniline (186 g, 2 mol) and ethanol (26 mL) was stirred at room temperature and 12M hydrochloric acid (167 mL) was added for 30 minutes. The dark yellow solution was refluxed at 85 ° C. for 7 days. The solution was cooled and diluted with ethyl acetate. The mixture was stirred in an ice bath and made alkaline (pH = 9) with 27% sodium hydroxide solution while maintaining the temperature below 30 ° C.

The organic layer was separated, washed with brine (3 ×), dried over magnesium sulfate and concentrated to give a brown oil (131 g). Excess of aniline was removed under reduced pressure. A catalytic amount of 12M hydrochloric acid (1 mL) was added and the residue was parted under high vacuum. The fraction distilled at 145 to 175 ° C. (0,2 mmHg) was collected and subjected to column chromatography (silica gel), eluting with 30% ethyl acetate toloene, 4- (2-methylcyclohex-1-enyl). A 9: 1 mixture (8.7 g) of aniline and (R, S) -4- (6-methylcyclohex-1-enyl) aniline was obtained.

4- (2-methylcyclohex-1-enyl) aniline:

¹ H NMR (DMSO- d ₆ ): δ 1,53 (s, 3H), 1,61 (m, 4H), 2,00 (bs, 2H), 2,13 (bs, 2H), 4,92 ( s, 2H), 6,50 (d, 2H), 6,79 (d, 2H); HPLC-MS (Method B): m / z = 188 (M ⁺ ); R _t = 2,96 min.

(R, S) -4- (6-methylchlorohex-1-enyl) aniline:

_{^{1 H NMR (DMSO- d 6)}} : 0,88 (d, 3H), 1,61 (m, 4H), 2,00 (bs, 2H), 2,13 (bs, 2H), 2,74 ( m, 1H), 4,92 (s, 2H), 5,68 (t, 1H), 6,50 (d, 2H), 6,79 (d, 2H).

Building Block 12: 4- (4-tert-butylcyclohex-1-enyl) aniline

4- (4-tert-butylcyclohex-1-enyl) aniline was prepared similarly to the preparation of building blocks using 4-tert-butylcyclohexanone (0.59 mol) and aniline.

_{^{1 H NMR (DMSO- d 6)}} : 0,88 (s, 9H), 1,21 (m, 2H), 1,90 (m, 2H), 2,10-2,50 (m, 3H), 4,97 (s, 2H), 5,90 (m, 1H), 6,50 (d, 2H), 7,06 (d, 2H); HPLC-MS (Method B): m / z = 230 (M ⁺ ); R _t = 4,07 min.

Building Blocks 13: (R, S) -4- (5-methylcyclohex-1-enyl) aniline and (R.S) -4- (3-methylcyclohex-1-enyl) aniline

A mixture of (R, S) -3-methylchlorohexanone (123 mL, 1.0 mol), aniline (182 mL, 2.0 mol), 12 M hydrochloric acid (167 mL, 2.0 mol), and ethanol (26 mL) at 10 ° C. Reflux for days. The solution was cooled and diluted with ethyl ester. The aqueous layer was made alkaline (pH = 10) using 6M sodium hydroxide solution. The organic layer was separated, washed with brine (3 ×), dried over magnesium sulfate and concentrated to give a yellow oil. Excess of aniline was removed under reduced pressure. A catalytic amount of 12M hydrochloric acid (1 mL) was added and the residue was parted under high vacuum. The distilled portion at 123-128 ° C. (0.15-0.20 mmHg) was collected to give 21.0 g of oil. ¹ H-NMR is 3 each of (R, S) -4- (5-methylcyclohex-1-enyl) aniline and (R, S) -4- (3-methylcyclohex-1-enyl) aniline 3: 2 indicates the presence of the mixture.

(R, S) -4- (5-methylcyclohex-1-enyl) aniline: ¹ H NMR (DMSO- d ₆ ): δ1.00 (d, 3H), 1.45-1.95 (m, 3H), 2.10 -2.45 (m, 3H), 5.04 (s, 2H), 5.86 (t, 1H), 6.48 (d, 2H), 7.06 (d, 2H).

(R, S) -4- (3-methylcyclohex-1-enyl) aniline: ¹ H NMR (DMSO- d ₆ ): δ1.00 (d, 3H), 1.45-1.95 (m, 3H), 2.10 -2.45 (m, 3H), 5. 04 (s, 2H), 5.75 (d, 1H), 6.48 (d, 2H), 7.06 (d, 2H).

General Method for Solution Phase Synthesis of Compounds of Formulas (Ia) and (Ib)

Wherein R ² , R ³ , R ⁷ , R ⁸ , A, E and D are as defined in formula (I).

Compounds made according to this general method (A) can be prepared via either the ester route or the carboxylic acid route. The only difference between these two pathways is the protection of benzoic acid as ester. Deprotection of the ester (step 2a) provides the same intermediate as that of the carboxylic acid pathway.

This method according to the ester route is described in Examples 1 and 2 and the method according to the carboxylic acid route is described in Example 3.

Example 1 (General Method (A))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (3-methoxy-5-trifluoromethylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

Step 1: 4-((4-cyclohexylphenylamino) methyl) benzoic acid methyl ester

4-formylbenzoic acid methyl ester (6.65 g, 40.5 mmol) was dissolved in hot methanol (175 mL). To this mixture, 4-cyclohexylaniline (7.1 g, 40.5 mmol) was added. To the resulting suspension, more methanol (75 mL) was added and the mixture was heated to reflux for 1 hour. After cooling to 0 ° C., the mixture was filtered to wash the solids with ice-cold methanol and dried in vacuo at 40 ° C. for 16 hours to give 10.95 g of 4-[(4-cyclohexylphenylimino) -methyl] benzoate An ester was obtained. This compound (10.93 g, 34 mmol) was suspended in methanol (200 mL), cold acetic acid (27 mL) was added, and sodium cyanoborohydride (1.9 g, 30 mmol) was added in portions. The mixture was stirred at rt for 1 h and concentrated in vacuo. The residue was dissolved in DCM (200 mL) and washed with 5% aqueous sodium carbonate (5 x 80 mL), then dried (magnesium sulfate) and concentrated in vacuo. Ethyl acetate (100 mL) and n-heptane (200 mL) were added to the residue and concentrated in vacuo to half the original volume. The solid was filtered, washed with n-heptane and dried in vacuo at 40 ° C. for 16 hours to yield 9.52 g (87%) of 4-((4-cyclo-hexylphenylamino) methyl) benzoic acid methyl ester.

¹ H-NMR (DMSO- d ₆ ): δ 1.2-1.4 (5H, m), 1.65 (5H, m), 2.30 (1H, t), 3.84 (3H, s), 4.30 (2H, d), 6.18 (1H, t), 6.50 (2H, d), 6.87 (2H, d), 7.49 (2H, d), 7.92 (2H, d); HPLC-MS (Method B): m / z = 324 (M + l); R _t = 7. 18 min.

Step 2: 4- [1- (cyclohexylphenyl) -3- (3-methoxy-5-trifluoromethylphenyl) ureidomethyl] benzoic acid methyl ester

5-methoxy-3- (trifluoromethyl) aniline (2.0 g, 10.5 mmol) was dissolved in ethyl acetate (10 mL), anhydrous HCl in ethyl ester (15 mL) was added and the solvent removed in vacuo. . The solid was co-evaporated with toluene (3 x 15 mL). Toluene (75 mL) and diphosgene (13 mL) were added and the reaction mixture was refluxed for 2.5 h under a nitrogen atmosphere. Excess diphosgene was removed in vacuo and the clear oil was co-evaporated with toluene. The obtained isocyanate was used without further purification

The above isocyanate was dissolved in DCM (75 mL) and 4-((4-cyclohexylphenylamino) -methyl) benzoic acid methyl ester (2.3 g, 7.1 mmol) was added. After stirring the reaction mixture overnight at room temperature, the solvent was removed in vacuo and the residual oil was purified by column chromatography on silica gel eluted with a mixture of heptane and ethyl acetate (7: 3). 3 g of 4- [1- (cyclohexylphenyl) -3- (3-methoxy-5-trifluoro-methylphenyl) ureidomethyl] benzoic acid methyl ester were obtained in oil form.

¹ H-NMR (DMSO-d6): δ1.22 (broad, 1H), 1.37 (broad, 4H), 1.7 (broad, 1H), 1.79 (broad, 4H), 3.77 (s, 3H), 3.83 (s , 3H), 4.98 (s, 2H), 6.81 (s, 1H), 7.18 (d, 2H), 7.23 (d, 2H), 7.42 (m, 3H), 7.51 (s, 1H), 7.90 (d, 2H), 8.53 (s, 1 H), 10.01 (s, 1 H); HPLC-MS (Method A): m / z = 541 (M + l); R _t = 8.98 min.

Step 2a: 4- [1- (Cyclohexylphenyl) -3- (3-methoxy-5-trifluoromethylphenyl) ureidomethyl] benzoic acid

4- [1- (cyclohexylphenyl) -3- (3-methoxy-5-trifluoromethylphenyl) ureidomethyl] benzoic acid methyl ester (3.0 g) is dissolved in anhydrous ethanol (50 mL) and sodium hydroxide (4N, 15 mL) was added and the reaction mixture was stirred for 16 h at room temperature. The organic solvent was removed in vacuo and additional water (50 mL) was added, then the pH was adjusted to the acidic reaction with hydrochloric acid (4N) and then ethyl acetate (200 mL) was added. The organic phase was washed with water (5 x 50 mL), dried (magnesium sulfate), filtered and evaporated in vacuo. The residue was recrystallized from acetonitrile (25 mL) to give 4- [1- (cyclohexylphenyl) -3- (3-methoxy-5-fluoromethylphenyl) ureidomethyl] benzoic acid (1.83 g) as crystals. Got it.

¹ H-NMR (DMSO- d ₆ ): δ1.22 (m, 1H), 1.37 (m, 4H), 1.70 (m, 1H), 1.79 (m, 4H), 3.77 (s, 3H), 4.95 ( s, 2H), 6.81 (s, 1H), 7.18 (d, 2H), 7.23 (d, 2H), 7.40 (d, 2H), 7.42 (s, 1H), 7.51 (s, 1H), 7.89 (d , 2H), 8.55 (s, 1 H), 12.90 (s, 1 H); HPLC-MS (Method A): m / z = 527 (M + l); R _t = 8.23 min; Mp 148-150 ° C.

Microanalysis theoretics for C ₂₉ H ₂₉ N ₂ F ₃ 0 ₄ :

C, 66.15%; H 5.55%; N 5.32%. Found:

C, 66.65%; H 5.70%; N 5.33%.

Step 3: (R) -3- {4- [1- (4-cyclohexylphenyl) -3- (3-methoxy-5-trifluoromethylphenyl) ureidomethyl] -benzoylamino} -2-hydrate Roxypropionic acid ethyl ester

4- [1- (cyclohexylphenyl) -3- (3-methoxy-5-trifluoromethylphenyl) ureidomethyl] benzoic acid (420 mg, 0.8 mmol) was dissolved in DMF (10 mL), followed by HOBt (160 mg, 1.2 mmol) and EDAC (230 mg, 1.2 mmol) were added. After the reaction was left for 30 minutes, (R) -isoserine ethyl ester (260 mg, 1.2 mmol) and diisopropylethylamine (210 mL, 1.2 mmol) dissolved in DMF (5 mL) were added and 16 at room temperature. The reaction mixture was stirred for hours. Water (50 mL) and ethyl acetate (100 mL) were added and the organic phase was washed with water (5 x 50 mL), then dried (magnesium sulfate), filtered and evaporated in vacuo. Purify the residue by column chromatography on silica gel, eluting with a mixture of heptane and ethyl acetate (1: 3) to give 510 mg of (R) -3- {4- [1- (4-cyclohexylphenyl)-. 3- (3-methoxy-5-trifluoromethylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid ethyl ester was obtained as an amorphous solid.

¹ H-NMR (DMSO- d ₆ ): δ1.12 (t, 3H), 1.22 (m, 1H), 1.37 (m, 4H), 1.70 (m, 1H), 1.79 (broad, 4H), 3.41 ( m, 1H), 3.52 (m, 1H), 3.77 (s, 3H), 4.06 (q, 2H), 4.21 (q, 1H), 4.95 (s, 2H), 5.68 (d, 1H), 6.81 ( s, 1H), 7.18 (d, 2H), 7.33 (d, 2H), 7.33 (d, 2H), 7.42 (s, 1H), 7.51 (s, 1H), 7.77 (d, 2H), 8.47 (t , 1H), 8.53 (s, 1H); HPLC-MS (Method A): m / z = 658 (M + l); R _t = 8.17 min.

Step 4:

(R) -3- {4- [l- (4-cyclohexylphenyl) -3- (3-methoxy-5-trifluoromethylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid ethyl ester Was dissolved in ethanol (15 mL) and sodium hydroxide (2N, 2 mL) was added. The reaction mixture was stirred for 60 minutes at room temperature. The ethanol was then removed in vacuo, then water (50 mL) was added and the pH adjusted to the acidic reaction with 4N hydrochloric acid. Filtration and washing with water (5 x 5 mL) and drying in vacuo gave 460 mg of the title compound as a crystallized solid.

¹ H-NMR (DMSO- d ₆ ): δ1.22 (m, 1H), 1.37 (m, 4H), 1.70 (m, 1H), 1.79 (broad, 4H), 3.37 (m, 1H), 3.51 ( m, 1H), 3.77 (s, 3H), 4.09 (t, 1H), 4.95 (s, 2H), 6.80 (s, 1H), 7.18 (d, 2H), 7.23 (d, 2H), 7.33 (d , 2H), 7.42 (s, 1H), 7.51 (s, 1H), 7.77 (d, 2H), 8.47 (t, 1H), 8.53 (s, 1H); HPLC-MS (Method A): m / z = 614 (M + l); R _t = 7.67 min.

Theoretical analysis for C ₃₂ H ₃₄ N ₃ F ₃ 0 ₆ (+ 1.25 H ₂ 0):

C, 60.42%; H, 5.78%; N, 6.61%. Found:

C, 60.25%; H, 5.55%; N, 6.50%.

Example 2 (General Method (A))

(R) -3- {4- [3- (3,5-Bis (trifluoromethyl) phenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

Step 2: 4- [3- (3,5-Bis (trifluoromethyl) phenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid

4-((4-cyclohexylphenylamino) methyl) benzoic acid methyl ester (2.38 g, 7.36 mmol) was dissolved in DCM (150 mL) and 3,5-bis (trifluoromethyl) phenyl isocyanate (1.36 mL, 8.10 mmol) ) Was added and the mixture was stirred at rt for 16 h. The reaction mixture was washed with water (3 x 15 mL), dried (magnesium sulfate) and concentrated in vacuo to give 4.3 g of 4- [3- (3,5-bis (trifluoromethyl) phenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid methyl ester was obtained.

¹ H-NMR (DMSO- d ₆ ): δ 1.17-1.44 (m, 5H), 1.66-1.82 (m, 5H), 3.83 (s, 3H), 4.98 (s, 2H, 7.207.28 (m, 4H), 7.44 (d, 2H), 7.62 (s, 1H), 7.93 (d, 2H), 8.24 (s, 2H), 8.94 (s, 1H); HPLC-MS (method A): m / z = 579 (M + 1); R _t = 9.50 min.

Step 2a: 4- [3- (3,5-Bis (trifluoromethyl) phenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid

4- [3- (3,5-Bis (trifluoromethyl) phenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid methyl ester (4.2 g, 7.36 mmol) in ethanol (80 mL) After suspension, sodium hydroxide (4N, 11 mL) was added and stirred for 16 h at room temperature. The reaction mixture was dried in vacuo and concentrated and acidified with hydrochloric acid (4 N, 12 mL) by addition of water (50 mL) to the residue. The aqueous phase was extracted twice with ethyl acetate (75 mL and 25 mL) and the combined organic phases were washed with water (3 x 15 mL), dried (magnesium sulfate) and concentrated in vacuo to afford 4- [3- (3 , 5-bis (trifluoromethyl) phenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid was obtained.

¹ H-NMR (DMSO- d ₆ ): δ1.32-1.43 (m, 5H), 1.7 (m, 1H), 1.75-1.85 (5H), 4.98 (s, 2H), 7.20-7.28 (m, 4H ), 7.4 (d, 2H), 7.61 (d, 1H), 7.88 (d, 2H), 8.25 (s, 2H), 8.93 (s, 1H), 12.90 (s, 1H); HPLC-MS (Method B): m / z = 565 (M + l); R _t = 8.65 min; M. p. 148.5-149.5 ° C.

Microanalysis theoretics for C ₂₉ H ₂₆ F ₆ N ₂ 0 ₃ :

C, 61.70%; H, 4.64%; N, 4.96%. Found:

C, 61.54%; H, 4.71%; N, 4.92%.

Step 3: (R) -3- {4- [3- (3,5-Bis (trifluoromethyl) phenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydrate Roxypropionic acid ethyl ester

4- [3- (3,5-Bis (trifluoromethyl) phenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid (0.22 g, 0.39 mmol) is dissolved in DMF (3 mL). 1-hydroxy-7-azabenzotriazole (0.06 g, 0.47 mmol) and EDAC (0.09 g, 1.2 mmol) were added. The mixture was stirred for 1.5 h and (R) -isoserine ethyl ester (0.10 g, 0.59 mmol) and diisopropylethylamine (0.10 mL, 0.59 mmol) in DMF (2 mL) were added. The mixture was stirred at rt for 16 h. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (25 mL). The aqueous phase was extracted with ethyl acetate (10 mL). The combined organic phases were washed with hydrochloric acid (0.2 N, 3 × 10 mL) and water: saturated sodium chloride (1: 1), then dried (magnesium sulfate) and concentrated in vacuo. Purify the residue by column chromatography (35 g silica gel) using ethyl acetate and n-heptane (6: 4) as eluent to afford 0.27 g of (R) -3- {4- [3- (3,5- bis (trifluoromethyl) phenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid ethyl ester was obtained.

¹ H-NMR (DMSO- d ₆ ): δ 1.14 (t, 3H), 1.19-1.42 (m, 5H), 1.67-1.85 (m, 5H), 3.38-3.48 (m, 1H), 3.49-3.57 (m, 1H), 4.08 (q, 2H), 4.20 (m, 1H), 4.97 (s, 2H), 5.67 (d, 1H), 7.20-7.27 (m, 4H), 7.36 (d, 2H ), 7.62 (s, 1 H), 7.75 (d, 2 H), 8.24 (s, 2 H), 8.48 (t, 1 H), 8.90 (s, 1 H); HPLC-MS (Method A): m / z = 680 (M + l); R _t = 8.42 min.

Step 4:

(R) -3- {4- [3- (3,5-bis (trifluoromethyl) phenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionate ethyl Ester (0.26 g, 0.38 mmol) was dissolved in ethanol (96%, 15 mL) and sodium hydroxide (4 N, 0.57 mL, 2.3 mmol) was added. After stirring for 1 h at 25 ° C., the mixture was evaporated in vacuo, and water (30 mL) was added to the residue and acidified with hydrochloric acid (4 N, 0.62 mL). The aqueous phase was extracted twice with ethyl acetate (25 mL and 10 mL) and the combined organic phases were washed with saturated sodium chloride: water (1: 1), dried (magnesium sulfate) and concentrated in vacuo to afford 0.21 g of the title compound . Got it.

₁ H-NMR (DMSO- d ₆ ): δ1.21-1.45 (m, 5H), 1.66-1.86 (m, 5H), 3.37-3.44 (m, 1H), 3.53-3.60 (m, 1H), 4.18 (t, 1H), 4.95 (s, 2H), 7.18-7.27 (m, 4H), 7.45 (d, 2H), 7.60 (s, 1H), 7.78 (d, 2H), 8.24 (s, 2H), 8.44 (t, 1 H), 8.90 (s, 1 H); HPLC-MS (Method B): m / z = 652 (M + l); R _t = 7. 93 min.

Example 3 (General Method (A))

(R) -3- {4- [3- (3-bromophenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

Step 1: 4-[(4-cyclohexylphenylamino) methyl] benzoic acid

4-cyclohexylaniline (8.0 g, 53 mmol) was dissolved in methanol (200 mL), then a suspension of 4-formylbenzoic acid (9.4 g, 53 mmol) in cold acetic acid (12 mL) was added in portions and the resulting mixture Was heated at reflux for 1.5 h. After cooling to room temperature sodium cyanoborohydride (5.0 g, 80 mmol) in methanol (100 mL) was added portionwise and the resulting mixture was stirred at room temperature for 16 hours. The mixture was filtered, washed thoroughly with water and dried at 50 ° C. for 3 days to give 12.8 g (78%) of 4-[(4-cyclohexylphenylamino) methyl] benzoic acid.

¹ H-NMR (DMSO- d ₆ ): 1.1-1.35 (5H, m), 1.65-1.75 (5H, m), 2.29 (1H, m), 4.31 (2H, s), 6.15 (1 H, bs) , 6.45 (2H, d), 6.89 (2H, d), 7.46 (2H, d), 7.88 (2H, d).

Step 2: 4- [3- (3-bromophenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid

3-bromoaniline (1.4 g, 8.1 mmol) was dissolved in diethyl ether (50 mL) and 3.5 M anhydrous HCI in ethyl acetate (2.3 mL) was added. The mixture was concentrated in vacuo. Toluene (100 mL) was added to the residue and concentrated in vacuo. Toluene (100 mL) and diphosgene (8.1 g, 41 mmol) were added to the residue and the resulting mixture was refluxed for 1.5 h. After cooling, the mixture was concentrated in vacuo. The residue was dissolved in toluene (100 mL) and concentrated in vacuo. The residue was dissolved in DMF (30 mL) and 4-[(4-cyclohexylphenylamino) methyl] benzoic acid (1.3 g, 4.1 mmol) was added. The mixture was stirred at rt for 16 h. Ethyl acetate (150 mL) was added to the mixture and washed with water: brine (1: 1) (2 x 100 mL). The organic phase was dried over sodium sulfate and concentrated in vacuo. First, the residue was purified by column chromatography on silica gel eluted with a mixture of ethyl acetate: n-heptane: triethylamine (7: 2: 1) and then with ethyl acetate and finally with methanol (1.95 g ( 94%) of 4- [3- (3-bromophenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid was obtained.

¹ H-NMR (CDCl ₃ ): δ 1.2-1.4 (5H, m), 1.7-1.8 (5H, m), 4.94 (2H, s), 7.1-7.25 (6H, m), 7.30 (2H, d ), 7.44 (1 H, d), 7.78 (1 H, t), 7.83 (2 H, d), 8.38 (1 H, s); HPLC-MS (Method B): m / z = 507 (M + l); R _t = 5.52 min.

Step 3: (R) -3- {4- [3- (3-bromophenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid ethyl ester

4- [3- (3-bromophenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid (0.20 g, 0.39 mmol) is dissolved in DMF (2.5 mL) and EDAC (0.12 g, 0.6 mmol) ) And HOBt (0.089 mg, 0.6 mmol) were added and then the mixture was stirred at room temperature for 10 minutes. (R) -isoserine ethyl ester hydrochloride (0.10 g, 0.6 mmol) and N, N-diisopropylethylamine (130 μL) dissolved in DMF (2.5 mL) were added and the resulting mixture was stirred at rt for 16 h. It was. To the mixture was added ethyl acetate (70 mL) and washed with water (2 x 100 mL), then the organic phase was dried over sodium sulfate and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with an ethyl acetate: n-heptane (1: 2) mixture containing 10% acetic acid. This resulted in 100 mg of (R) -3- {4- [3- (3-bromophenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid ethyl ester.

HPLC-MS (Method B): m / z = 624 (M + l); R _t = 5.33 min.

Step 4: (R) -3- {4- [3- (3-bromophenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

(R) -3- {4- [3- (3-bromophenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino-2-hydroxypropionic acid ethyl ester (100 mg) 10 mL), 1 N sodium hydroxide (480 μL) was added and the resulting mixture was stirred at rt for 1 h. 1 N hydrochloric acid (480 μL) was added and the mixture was concentrated in vacuo. The residue was suspended in water (50 mL) and filtered to give 38 mg of the title compound .

¹ H-NMR (CDCl ₃ ): δ 1.2-1.4 (5H, m), 1.7-1.85 (5H, m). 2.45 (1H, s), 3.7 (2H, m), 4.30 (1H, m), 4.78 (2H, s), 6.23 (1 H, s), 6.95-7.3 (8H, m), 7.42 (1 H, s), 7.60 (2 H, d); HPLC-MS (Method A): m / z = 595 (M + l); R _t = 7.48 min.

Example 4 (General Method (A))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (4-trifluoromethylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

¹ H-NMR (DMSO- d ₆ ): δ1.2-1.4 water H, m), 1.7-1.8 (5H, m), 3.40 (1H, m), 3.56 (1H, dt), 4.18 (1H, t ), 4.97 (2H, s), 5.5 (1H, brd) 7.14-7.25, (6H, m), 7.34 (2H, d), 7.55 (2H, d), 7.79 (2H, d), 8.38 (1H, s), 8.44 (1 H, t); HPLC-MS (Method A): m / z = 600 (M + l); R _t = 7.38 min.

Example 5 (General Method (A))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (3-trifluoromethylphenyl) ureidomethyl] benzoylamino-2-hydroxypropionic acid

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 1.15-1-45 (m, 5H), 1.60-1.90 (m, 5H), 4.95 (s, 2H), 7.12 (d, 2H), 7.19 (d, 2H), 7.24 (d, 1H), 7.45 (t, 1H), 7.72 (d, 2H), 7.75 (s br, 1H), 7.90 (s, 1H), 8.55 (s, 1H), 8.58 (s br, 1 H); HPLC-MS (Method B): m / z = 584 (M + l); R _t = 4.99 min.

Example 6 (General Method (A))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (3-fluoro-5-trifluoromethylphenyl) ureido] benzoylamino} -2-hydroxypropionic acid

¹ H-NMR (200 MHz, DMSO- d ₆ ): δ1. 15-1-45 (m, 5H), 1.60-1.85 (m, 5H), 3.25-3.65 (m, 3H), 4.13 (t, 1H), 4.95 (s, 2H), 7.10-7.22 (m, 5H ), 7.30 (d, 2H), 7.75 (m, 3H), 8.45 (t, 1H), 8.78 (s, 1H); HPLC-MS (Method A): m / z = 602 (M + l); R _t = 7.83 min.

Example 7 (General Method (A))

(R) -3- {4- [3- (3-cyano-5-trifluoromethylphenyl) -1- (4-cyclohex-1-enylphenyl) ureidomethyl] benzoylamino} -2-hydrate Roxypropionic acid

¹ H-NMR (200 MHz, DMSO- d ₆ ): δ 1.15-1.70 (m, 4H), 2.12 (s, 2H), 2.35 (s, 2H), 5.00 (s, 2H), 6.15 (s, 1H), 7.10-7.40 (m, 6H), 7.78 (m, 3H), 8.20 (d, 2H), 8.90 (s, 1H); HPLC-MS (Method A): m / z = 607 (M + l); R _t = 7.42 min.

Example 8 (General Method (A))

(R) -3- {4- [3- (3-cyano-5-trifluoromethylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 1.15-1.45 (m, 5H), 1.60-1.85 (m, 5), 3.45-3.60 (m, 3H), 4.15 (t, 1H) 4.95 (s, 2H), 7.20 (dd, 4H), 7.35 (d, 2H), 7.76 (d, 2H), 7.88 (s, 1H), 8.18 (s, 1H), 8.21 (s, 1H), 8.45 (t, 1H), 8.95 (s, 1H); HPLC-MS (Method A): m / z = 609 (M + l); R _t = 7.58 min.

Example 9 (General Method (A))

(R) -3- {4- [3- (3-bromo-5-trifluoromethylphenyl) -1- (4-cyclohex-1-enylphenyl) ureidomethyl] benzoylamino} -2-hydrate Roxypropionic acid

_{^{1 H-NMR (DMSO- d 6}} ): δ1. 50-1.76 (m, 4H), 2.15 (m, 2H), 2.33 (m, 2H), 3.37 (m, 2H), 3.54 (m, 1H), 4.14 (dd, 1H), 4.95 (s, 2H) , 6.17 (t, 1H), 7.17 (d, 2H), 7.33 (d, 2H), 7.40 (d, 2H), 7.46 (s, 1 H), 7.75 (d, 2H), 7.91 (s, 1 H ), 8.07 (s, 1 H), 8.44 (t, 1 H), 8.66 (s, 1 H).

Example 10 (General Method (A))

(R) -3- {4- [1- (4-cyclohex-1-enylphenyl) -3- (3-methoxy-5-trifluoromethylphenyl) ureidomethyl] benzoylamino} -2-hydrate Roxypropionic acid

_{^{1 H-NMR (DMSO- d 6}} ): δ1. 51-1. 76 (m, 4H), 2.15 (m, 2H), 2.34 (m, 2H), 3.37 (m, 2H), 3.54 (m, 1H), 3.75 (s, 3H), 4.14 (dd, 1H), 4.95 (s, 2H), 6.16 (t, 3H), 6.78 (s, 1H), 7.16 (d, 2H), 7.32 (d, 2H), 7.38 (d, 2H), 7.43 (s, 1H), 7.76 ( d, 2H), 8.44 (t, 1 H), 8.51 (s, 1 H).

Example 11 (General Method (A))

(R) -3- {4- [3- (3-bromophenyl) -1- (4-cyclohex-1-enylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

The title compound was prepared according to General Method (A), in which Step 2 was modified as follows:

Step 2: 4- [3- (3-bromophenyl) -l- (4-cyclohex-l-enylphenyl) ureidomethyl] benzoic acid methyl ester

To 3-bromobenzoic acid (1.2 g, 5.7 mmol) in toluene (20 mL) add triethylamine (0.91 mL, 6.5 mmol) and diphenoxyphosphoryl azide (1.3 mL, 6.2 mmol) and stir the mixture Heated to 100 ° C. After 1 hour 4-[(4-cyclohex-1-enylphenylamino) methyl] benzoic acid methyl ester (prepared similar to that described in Example 1, step 1) (1.6 g, 5 mmol) was added and additional Heating was continued for 1.5 hours. After cooling to room temperature the mixture was transferred to a separatory funnel with ethyl ester (50 mL). The organic mixture was washed with saturated aqueous sodium hydrogen carbonate (2 x 50 mL) and back washed with ethyl acetate (50 mL). The organic phase was collected, dried (sodium sulfate) and the solvent was removed in vacuo to yield a brown oil which was purified on silica gel eluted with DCM to give 700 mg of 4- [3- (3-bromophenyl) -1- (4-cyclohex-1-enylphenyl) ureidomethyl] benzoic acid methyl ester was obtained.

HPLC-MS (Method B): m / z = 521 (M + l); R _t = 6.1 min.

Data for the title compound :

¹ H-NMR (CDCl ₃ ): δ1.54 (s, 2H), 1.65 (s, 2H), 2.15 (s, 2H), 2.36 (s, 2H), 3.56 (br s, 2H), 4.19 (br s, 1H), 4.71 (s, 2H), 6.09 (s, 1H), 6.42 (s, 1H), 6.90-7.18 (m, 6H), 7.30.7.70 (m, 6H); HPLC-MS (Method B): m / z = 592.5 (M + l); R _t = 4.73 min.

Example 12 (General Method (A))

(R) -3- {4- [3- (3-bromo-5-trifluoromethylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

¹ H-NMR (DMSO- d ₆ ): δ 1.14-1.43 (m, 5H), 1.64-1.83 (m, 5H), 3.40-3.45 (m, 1H), 3.45-3.53 (m, 1H), 4.00 (t, 1H), 4.95 (s, 2H), 7.13-7.27 (m, 4H), 7.33 (d, 2H), 7.48 (s, 1H), 7.77 (d, 2H), 7.92 (s, 1H) , 8.07 (s, 1 H), 8.43 (t, 1 H), 8.71 (s, 1 H); HPLC-MS (Method A): m / z = 662 (M + l); R _t = 8.17 min.

Example 13 (General Method (A))

(S) -trans-3- {4- [3- (3,5-Bis (trifluoromethyl) phenyl) -1- (4-tert-butylcyclohexyl) ureidomethyl] benzoylamino} -2- Hydroxypropionic acid

Step 1: trans-4-[(4-tert-butylcyclohexylamino) methyl] benzoic acid methyl ester

4-formibenzoic acid methyl ester (10.6 g, 64.4 mmol) is dissolved in methanol (200 mL). A 17:83 cis / trans mixture of 4-tert-butylcyclohexylamine (10.0 g, 64.4 mmol, Aldrich) was added, resulting in immediate precipitation of white nodules. The mixture was heated to reflux for 30 minutes to complete imine formation, then cooled to 0 ° C. in an ice bath. Crystalline pure trans-4-[(4-tert-butylcyclohexylimino) methyl] benzoic acid methyl ester was collected by filtration and dried in vacuo overnight. Yield: 15.3 g (78%).

₁ H NMR (CDCl ₃ ), 300 MHz: δ 8.37 (s, 1H); 8.06 (d, 2 H); 7.77 (d, 2 H); 3.92 (s, 3 H); 3.17 (m, 1 H); 1.83 (m, 4 H); 1.60 (m, 2 H), 1.09 (m, 3 H); 0.87 (s, 9 H).

Theoretical Analysis for C ₁₉ H ₂₇ NO ₂

C, 75. 71%; H, 9.03%; N, 4.65%. Found:

C, 75.60%; H, 9.37%; N, 4.68%.

The mother liquor was dried to give 4.2 g (22%) of a white solid, which consists mainly of the imino cis isomer according to NMR.

¹ H NMR (CDCl ₃ ), 300 MHz: δ 8.36 (s, 1H); 8.07 (d, 2 H); 7.81 (d, 2 H); 3.92 (s, 3 H); 3.54 (m, 1 H); 1.55-1.92 (m, 8 H); 1.14 (m, 1 H); 0.90 (s, 9 H).

Trans-4-[(4-tert-butylcyclohexylimino) methyl benzoic acid methyl ester (21.0 g, 69.2 mmol) was suspended in methanol (300 mL) and acetic acid (50 mL) was added. To the resulting clear solution sodium cyanoborohydride (3.5 g, 55.5 mmol) was added and the mixture was stirred at ambient temperature for 30 minutes. The reaction volume was reduced to 1/3 by rotary evaporation and ethyl acetate (500 mL) was added. The organic phase was washed with sodium carbonate solution (5%, 500 mL) and dried over sodium sulfate. The solvent was removed by rotary evaporation to yield the title material in solid form of white crystalline pure enough for further reaction. Yield: 21 1 g (100%).

¹ H NMR (CDCl ₃ ), 300 MHz: δ 7.98 (d, 2H); 7.38 (d, 2 H); 3.90 (s, 3 H); 3.86 (s, 2 H); 2.39 (m, 1 H); 2.01 (m, 2 H); 1.77 (m, 2 H); 1.51 (bs, 1 H); 0.93-1.18 (m, 5 H); 0.82 (s, 9 H); HPLC-MS (method _{B): R t = 4.87 m} / z = 304 (M + 1).

Step 2: trans-4- [l- (3,5-bis (trifluoromethyl) phenyl) -3- (4-tert-butylcyclohexyl) ureidomethyl] benzoic acid methyl ester

Trans-4-[(4-tert-butylcyclohexylamino) methyl] benzoic acid methyl ester (1.0 g, 3.3 mmol) was dissolved in acetonitrile (40 mL) and then 3,5-bis (trifluoromethyl) Phenyl isocyanate (0.9 g, 3.6 mmol) was added and the reaction stirred overnight at room temperature. Concentrate the solvent in vacuo to 5-10 mL and separate the crystals by filtration to yield 1.45 g (81%) of trans-4- [1- (3,5-bis (trifluoromethyl) phenyl) -3- ( 4-tert-butylcyclohexyl) ureidomethyl] benzoic acid methyl ester was obtained.

_{^{1 H-NMR (DMSO- d 6}} ): δ9.08 (s, 1H); 8.25 (s, 2 H); 7.91 (d, 2 H); 7.60 (s, 1 H); 7.40 (d, 2 H); 4.65 (s, 2 H); 4.07 (broad, 1 H); 3.83 (s, 3 H); 1.71 (broad, 4 H); 1.42 (broad, 2 H); 1.11 (broad, 2 H); 0.93 (broad, 1 H); HPLC-MS (Method B): m / z = 559 (M + l); R _t = 9.40 min; M. p. 188-190 ° C. (CH ₃ CN).

Theoretical analysis for C ₂₈ H ₃₂ N ₂ F ₆ 0 ₃ :

C, 60.21%; H, 5.77%; N, 5.02%. Found:

C, 60.46%; H, 5.94%; N, 5.00%.

Step 2a: trans-4- [1- (3,5-bis (trifluoromethyl) phenyl-3- (4-tert-butylcyclohexyl) ureidomethyl] benzoic acid

Trans-4- [l- (3,5-bis (trifluoromethyl) phenyl) -3- (4-tert-butylcyclohexyl) ureidomethyl] benzoic acid methyl ester (1.4 g) was dissolved in anhydrous ethanol (20 mL ), Sodium hydroxide (2N, 11 mL) was added and the reaction mixture was gently refluxed for 2 hours. The ethanol was removed in vacuo, additional water (40 mL) was added, the pH was adjusted to the acidic reaction with hydrochloric acid (4 N) and then ethyl acetate (200 mL) was added. The organic phase was separated, washed with water (4 x 50 mL), dried over magnesium sulfate, filtered and evaporated in vacuo to give 1.1 g (85%) trans-4- [1- (3,5-bis ( Trifluoromethyl) phenyl) -3- (4-tert-butylcyclohexyl) ureidomethyl] benzoic acid was obtained in solid form.

_{^{1 H-NMR (DMSO- d 6}} ): δ12.85 (s, 1H); 9.08 (s, 1 H); 8. 25 (s, 2 H); 7.89 (d, 2 H); 7.61 (s, 1 H); 7.38 (d, 2 H); 4.65 (s, 2 H); 4.07 (m, 1 H); 1.73 (m, 4 H); 1.43 (m, 2 H); 1.11 (m, 2 H); 0.93 (m, 1 H); 0.82 (s, 9 H); M. p. 239-241 ° C. (MeCN).

Microanalytic Theory for C ₂₇ H ₃₀ N ₂ F ₆ 0 ₃ :

C, 59.55%; H, 5.55%; N, 5.14%. Found:

C, 59.58%; H, 5.65%; N, 5.11%.

The next steps 3 and 4 gave the title compound .

¹ H-NMR (DMSO- d ₆ ): δ0.83 (s, 9H), 0.90-0.99 (m, 1H), 1.06-1.15 (m, 2H), 1.37-1.50 (m, 2H), 1.64-1.80 (m, 4H), 3.35-3.43 (m, 1H), 3.52-3.61 (m, 1H), 4.01-4.11 (m, 1H), 4.18 (t, 1H), 4.63 (s, 2H), 7.35 (d , 2H), 7.61 (s, 1H), 7.81 (d, 2H), 8.27 (s, 2H), 8.43 (t, 1H), 9.07 (s, 1H), 12.46 (broad, 1H); HPLC-MS (Method A): m / z = 632 (M + l); R _t = 8.00 min; M. p. 185-187 ° C.

Theoretical analysis for C ₃₀ H ₃₅ F ₆ N ₃ 0 ₅ :

C, 57.05%; H, 5.59%; N, 6.65%. Found:

C, 57.32%; H, 5.70%; N, 6.27%.

Example 14 (General Method (A))

(R) -trans-3- {4- [3- (3,5-bis (trifluoromethyl) phenyl) -1- (4-tert-butylcyclohexyl) ureidomethyl benzoylamino} -2-hydrate Roxypropionic acid

¹ H-NMR (DMSO- d ₆ ): δ0.82 (s, 9H), 0.93 (m, 1H), 1.11 (m, 2H), 1.43 (m, 2H), 1.71 (m, 4H), 3.38 ( m, 1H), 3.56 (m, 1H), 4.05 (m, 1H), 4.16 (t, 1H), 4.63 (s, 2H), 7.33 (d, 2H), 7.61 (s, 1H), 7.80 (d , 2H), 8.26 (s, 2H), 8. 43 (t, 1H); HPLC-MS (Method A): m / z = 632 (M + l); R _t = 8.17 min; Mp 184-187 캜.

Microanalytic Theory for C ₃₀ H ₃₅ N ₃ F ₆ 0 ₅ (+ 0.5 Ethyl Acetate):

C, 56.88%; H, 5.82%; N, 6.22%. Found:

C, 56.63%; H, 5.66%; N, 6.47%.

Example 15 (General Method (A))

Trans- (R) -3- {4- [3- (3-methyl-5-trifluoromethylphenyl) -1- (4-tert-butylcyclohexyl) ureidomethyl] benzoylamino} -2-hydroxy Propionic acid

_{^{1 H-NMR (DMSO- d 6}} ): δ0. 82 (s, 9H), 0.93 (m, 1H), 1.11 (m, 2H), 1.43 (m, 2H), 1.71 (m, 4H), 2.33 (s, 3H), 3.38 (m, 1H), 3.56 (m, 1H), 4.05 (m, 1H), 4.16 (t, 1H), 4.63 (s, 2H), 7.10 (s, 1H), 7.33 (d, 2H), 7.73 (s, 1H), 7.80 (d, 2H), 8.43 (t, 1H), 8.62 (s, 1H); HPLC-MS (Method A): m / z = 578 (M + l); R _t = 7. 45 min.

Example 16 (General Method (A))

(RS) -3- {4- [1- (4-tert-butylphenyl) -3- (4-trifluoromethoxyphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

Step 1: 4- [1- (4-tert-butylphenyl) -3- (4-trifluoromethoxyphenyl) ureidomethyl] benzoic acid

4-formibenzoic acid methyl ester (10.6 g, 64 mmol) was dissolved in methanol (200 mL). 4-tert-butylaniline (9.61 g, 64 mmol) was added and the resulting suspension was refluxed for 15 minutes. After cooling to silol, TFA (5.18 mL, 68 mmol) was added and sodium cyanoborohydride (3.26 g, 52 mmol) was added in portions. The resulting mixture was refluxed at room temperature for 2 hours and concentrated. The residue was partitioned between ethyl ester (200 mL) and 1N aqueous sodium hydroxide (150 and 100 mL). The organic phase was dried (magnesium sulfate) and evaporated in vacuo to afford 19.0 g (99%) of 4-[(4-tert-butylphenylamino) methyl] benzoic acid methyl ester in solid form.

¹ H-NMR (CDCl ₃ ): δ 1.28 (9H, s), 3.92 (3H, s), 4.39 (2H, s), 6.57 (2H, d), 7.20 (2H, d), 7.44 (2H, d), 8.00 (2H, d).

Step 2:

The above benzoic acid methyl ester (0.73 g, 2.44 mmol) was dissolved in acetonitrile (7 mL) and 4-trifluoromethoxyphenylisocyanate (405 μL, 2.68 mmol) was added. The resulting mixture was stirred at rt for 3 h and then refluxed for 1.5 h. After cooling and concentration in vacuo, the residue was purified first by column chromatography on silica gel, eluting first with ethyl acetate and heptane (1: 6), then eluting with ethyl acetate and heptane (1: 3). 1.14 g (94%) of 4- [1- (4-tert-butylphenyl) -3- (4-trifluoromethoxyphenyl) ureidomethyl] benzoic acid methyl ester was obtained in the form of an oil.

¹ H-NMR (CDCl ₃ ): δ 1.35 (9H, s), 3.91 (3H, s), 4.97 (2H, s), 6.30 (1H, s), 7.1 (4H, m), 7.327.43 ( 6H, m), 7.96 (2H, d). TLC: Rf = 0.11 (Si02; ethyl acetate / heptane (1: 6)); HPLC-MS (Method B): m / z = 501 (M + l); R _t = 9.05 min.

Step 2a:

The above ureidomethylbenzoic acid methyl ester (1.14 g, 2.28 mmol) was dissolved in 1,4-dioxane (25 mL) and 1N aqueous sodium hydroxide (5 mL) was added. The resulting mixture was stirred at rt for 1 h. Ethanol (15 mL) and 1 N aqueous sodium hydroxide (5 mL) were added and the resulting mixture was stirred at rt for 16 h. The mixture was concentrated in vacuo and partitioned between 1 N hydrochloric acid (100 mL) and ethyl acetate (2 × 50 mL). The combined organic phases were dried (magnesium sulfate) and concentrated in vacuo to give 847 mg (76%) of 4- [1- (4-tert-butylphenyl) -3- (4-trifluoromethoxyphenyl) ureidomethyl] Benzoic acid was obtained in solid form.

¹ H-NMR (CDCl ₃ ): δ1.33 (9H, s), 3.91 (3H, s), 4.97 (2H, s), 6.30 (1 H, s), 7.1 (4H, m), 7.33 (2H , d), 7.43 (4H, m), 8.03 (2H, d).

Step 3: (RS) -3- {4- [1- (4-tert-butylphenyl) -3- (4-trifluoromethoxyphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid ethyl ester Porn

To a solution of [1- (4-tert-butylphenyl) -3- (4-trifluoromethoxyphenyl) ureidomethyl] benzoic acid (1.0 g, 2.06 mmol) in DMF (12 mL) and DCM (10 mL). 1-hydroxy-7-azabenzo-triazole (0.33 g, 2.47 mmol) was added. After stirring for 1 hour at room temperature, EDAC (0.47 g, 2.47 mmol), (RS) -isoserine ethyl ester hydrochloride (0.52 g, 3.09 mmol) and diisopropylethylamine (1.1 mL, 6.18 mmol) were successively Was added. After stirring for 17 h at ambient temperature, the reaction mixture was partitioned between water, brine and ethyl acetate. The aqueous phase was further extracted with ethyl acetate. The combined organic phases were washed with water and brine. After drying (magnesium sulfate) and filtration, the organic phase was evaporated in vacuo. The residue was purified by column chromatography on silica gel eluted with a mixture of toluene and ethyl acetate (6: 4). This results in 1.2 g (97%) of (RS) -3-4- [1- (4-tert-butylphenyl) -3- (4-trifluoromethoxyphenyl) ureidomethyl] benzoylamino} -2-hydrate Roxypropionic acid ethyl ester was obtained in oil form.

¹ H-NMR (DMSO- d ₆ ): δ1.12 (t, 3H), 1.27 (s, 9H), 3.45 (m, 2H), 4.05 (q, 2H), 4.21 (dd, 1H), 4.98 ( s, 2H), 5.67 (d, 1H), 7.20 (dd, 4H), 7.35 (dd, 4H), 7.55 (d, 2H), 5.77 (d, 2H), 8.42 (s, 1H), 8.48 (t , 1H); HPLC-MS (Method B): m / z = 602 (M + l); R _t = 3.38 min.

Step 4:

A solution of 3-4- [1- (4-tert-butylphenyl) -3- (4-trifluoromethoxyphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid ethyl ester was dissolved in anhydrous ethanol (20 mL ) And 1M sodium hydroxide (6 mL) was added. Stirring was continued for 17 hours and the solution was acidified with aqueous hydrochloric acid. The solvent was allowed to stir and the remaining oil dissolved by applying heat to acetonitrile (20 mL). Water (20 mL) was added dropwise under vigorous stirring and the mixture was allowed to cool to room temperature. The precipitate was filtered off, washed with water and dried to afford 0.51 g (43%) of the title compound in solid form.

¹ H-NMR (DMSO- d ₆ ): δ 1.25 (s, 9H), 3.50 (ddd, 2H), 4.18 (dd, 1H), 4.95 (s, 2H), 7.20 (dd, 4H), 7.39 ( dd, 4H), 7.52 (d, 2H), 7.78 (d, 2H), 8.32 (s, 1H), 8.46 (t, 1H); HPLC-MS (Method B): m / z = 574 (M + l); R _t = 3.07 min.

Microanalytic Theory for C ₂₉ H ₃₀ F ₃ N ₃ O ₆ :

C, 60. 73%; H, 5. 27%; N, 7.33%. Found:

C, 60.77%; H, 5. 37; N%, 7.26%.

Example 17 (General Method (A)

(RS) -3- {4- [1-4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

Step 3: (RS) -3- {4- {1-4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2-hydroxypropionate ethyl ester

Yield: 0.33 g (89%).

¹ H-NMR (CDCl ₃ ): δ 1.30 (t, 3H), 1.68 (m, 2H), 1.79 (m, 2H), 2.23 (m, 2H), 2.38 (m, 2H), 3.41 (d, 1H), 3.83 (m, 2H), 4.27 (q, 2H), 4.37 (dd, 1H), 4. 92 (s, 2H), 6.20 (m, 1H), 6.27 (s, 1H), 6.51 (t , 1H), 6.98 (s, 1H), 7.04 (d, 2H), 7.18 (d, 2H), 7.33 (d, 2H), 7.42 (d, 2H), 7.68 (d, 2H).

Step 4: (RS) -3- {4- {1-4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

(RS) -3- {4- {1-4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoyl in ethanol (15 mL) and THF (15 mL) A solution of amino} -2-hydroxypropionic acid ethyl ester (0.99 g, 1.61 mmol) was stirred and 1 M sodium hydroxide (6 mL) was added. The mixture was stirred at 40 ° C. for 4 h and acidified with 1 N hydrochloric acid. After evaporation in vacuo the residue was purified on semipreparative HPLC (Gilson system). Pure portions were combined and evaporated in vacuo to give 0.74 g (79%) of the title compound in solid form.

¹ H-NMR (DMSO- d ₆ ): δ1.56 (m, 2H), 1.70 (m, 2H), 2.17 (s, 2H), 2.33 (s, 2H), 3.35 (m, 2H), 3.55 ( m, 2H), 4.15 (dd, 1H), 4.95 (s, 2H), 6.20 (s. 1H), 7.12 (s, 1H), 7.18 (d, 2H), 7.33 (d, 2H), 7.40 (d , 2H), 7.62 (s, 2H), 7.76 (d, 2H), 8.43 (t, 1H), 8.55 (s, 1H); HPLC-MS (Method B): m / z = 582 (M + l); R _t = 5.13 min.

Microanalytic Theory for C ₂₉ H ₃₀ Cl ₂ N ₃ 0 ₅ :

C, 61.86%; H, 5.02%; N, 7.21%. Found:

C, 61. 10%; H, 5.05%; N, 7.03%.

Example 18

(S) -3- {4- [l- (4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

4- [1- (4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoic acid (130 mg, 0.26 mmol) is dissolved in DMF (2 mL) and EDAC (50 mg, 0.26 mmol) and HOBt (43 mg, 0.32 mmol) were then added and the reaction mixture was stirred at rt for 1 h. The above crude (S) -2,2-dimethyl-5-oxo- [1,3] dioxolan-4-ylmethylammonium trifluoroacetate was dissolved in DMF (1 mL) and diisopropylethylamine (450 mg, 3.5 mmol) was added to the reaction mixture. The mixture was stirred at rt for 16 h. The reaction mixture was transferred to a silica gel column and eluted with DCM and evaporated to give crude (S) -4- [1- (4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl ] -N- (2,2-dimethyl-5-oxo- [1,3] dioxolan-4-ylmethyl) benzamide was obtained in oil form. The oil was dissolved in acetonitrile (5 mL), then hydrochloric acid (1 N, 5 mL) was added and the mixture was stirred at rt for 1.5 h. Solvent was evaporated off and the crude product was purified by semipreparative HPLC (acetonitrile / water gradient) to afford the title compound .

HPLC-MS (Method B): m / z = 582 (M + l); R _t = 5.10 min.

Example 19 (General Method (A))

(R) -3- {4- [1- (4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

Step 3: (R) -3- {4- [1- (4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid Ethyl ester

¹ H-NMR (acetone- d ₆ ): δ 1.20 (t, 3H), 1.70 (dm, 4H), 2.31 (dm, 4H), 3.70 (m, 2H), 4.15 (q, 2H), 4.35 ( dd, 1H), 5.02 (s, 2H), 6.22 (m, 1H), 7.00 (s, 1H), 7.20 (d, 2H), 7.40 (dd, 4H), 7.61 (ds, 2H), 7.80 (d , 3H), 7.89 (s, 1 H).

Step 4:

(R) -3- {4- [1- (4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl in ethanol (5 mL) and THF (5 mL)] Benzoylamino} -2-hydroxypropionic acid ethyl ester (0.60 g, 0.98 mmol) was stirred and 4N sodium hydroxide (0.76 mL, 2.94 mmol) was added. The solution was stirred for 3 hours at room temperature and acidified with 1N hydrochloric acid. Evaporation in vacuo gave an oil which was partitioned between ethyl acetate, water and brine. The aqueous phase was extracted with ethyl acetate and the combined organic phases were washed with water and brine. Drying (magnesium sulfate), filtration and evaporation in vacuo yielded 0.43 g (73%) of the title compound in solid form.

_{^{1 H-NMR (DMSO- d 6}} ): δ1. 50-1.80 (4H, m), 2.08-2.38 (4H, m), 3.36-3.65 (2H, m), 4.14-4.24 (1H, m), 4.96 (2H, m), 6.17 (1H, t), 7.14 (1H, t), 7.18 (2H, d), 7.35 (2H, d), 7.42 (2H, d), 7.63 (2H, d), 7.78 (2H, d), 8. 48 (1 H, t ), 8.55 (1 H, s); HPLC-MS (Method B): m / z = 582 (M + l); R _t = 5.11 min.

Example 20 (General Method (A))

(R) -3- {4- [3- (3-chlorophenyl) -1- (4-cyclohex-1-enylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

¹ H-NMR (DMSO- d ₆ ): δ1.56-1.71 (m, 4H), 2.15-2.33 (m, 4H), 3.37-3.51 (m, 2H), 4.14-4.17 (m, 1H), 4.96 (s, 1H), 6.17 (t, 1H), 7.12 (dd, 1H), 7.27-7.41 (m, 8H), 7.62 (t, 1H), 7.87 (d, 2H), 8.38-8.43 (m, 3H ); HPLC-MS (Method B): m / z = 548 (M + l); R _t = 4.69 min.

Example 21 (General Method (A))

(R) -3- {4- [1- (4-cyclohex-1-enylphenyl) -3-phenylureidomethyl] benzoylamino} -2-hydroxypropionic acid

¹ H-NMR (DMSO- d ₆ ): δ1.56-1.71 (m, 4H), 2.15-2.40 (m, 4H), 3.30-3.51 (m, 2H), 4.14-4.19 (m, 1H), 4.97 (s, 1H), 6.16 (t, 1H), 6.98 (t, 1H), 7.12-7.48 (m, 10H), 7.79 (d, 2H), 8.15 (s, 1H), 8.43 (t, 1H).

Example 22 (General Method (A))

(R) -3- {4- [3-benzyl-1- (4-cyclohex-1-enylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

¹ H-NMR (DMSO- d ₆ ): δ1.50-1.75 (m, 4H), 1.86 (s, 2H), 2.08 (s, 2H), 3.30-3.60 (m, 2H), 4.134.20 (m , 1H), 4. 25 (d, 2H), 4.87 (s, 2H), 6.18 (t, 1H), 6.55 (t, 1H), 7. 08-7.42 (m, 11H), 7.77 (d , 2H), 8.48 (t, 1H).

Example 23 (General Method (A))

(RS) -3- {4- [1- (4-cyclohex-1-enylphenyl) 3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2-fluoropropionic acid

Steps 1 and 2: 4- [1- (4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoic acid

¹ H-NMR (DMSO- d ₆ ): δ1.52-1.77 (m, 4H), 2.10-2.23 (m, 2H), 2.26-2.38 (m, 2H), 4.95 (s, 2H), 6.18 (t , 1H), 7.14 (t, 1H), 7.17 (d, 2H), 7.34 (d, 2H), 7.40 (d, 2H), 7.64 (dd, 2H), 7.85 (d, 2H), 8.55 (s, 1H).

Microanalytic Theory for C ₂₇ H ₂₄ N ₂ O ₃ Cl ₂ :

C, 65.46%; H, 4.88%; N, 5.65%. Found:

C, 65.43%; H, 5.10%; N, 5.66%.

Step 3: (RS) -3- {4- [1- (4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2-fluoropropionic acid Methyl ester

¹ H-NMR (DMSO- d ₆ ): δ1.52-1.75 (m, 4H), 2.10-2.40 (m, 4H), 3.60-3.81 (m, 5H), 4.95 (s, 2H), 5.04-5.35 (m, 1H), 6.18 (m, 1H), 7.10-7.80 (m, 11H), 8.55 (s, 1H), 8.75 (t, 1H), 13.45 (br s, 1H).

Step 4:

(RS) -3-4- [1- (4-cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2-fluoro in a mixture of THF and methanol Hydroropyonic acid methyl ester was hydrolyzed to afford the title compound.

¹ H-NMR (DMSO- d ₆ ): δ 1.59-1.72 (m, 4H), 2.15-2.33 (m, 4H), 3.58-3.81 (m, 2H), 4.96 (s, 2H), 5.17-5.23 (m, 1H), 6.18 (m, 1H), 7.13-7.80 (m, 11H), 8.54 (s, 1H), 8.73 (t, 1H), 13.45 (brs, 1H).

Example 24 (General Method (A))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (4-trifluoromethylsulfanylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

Step 2: 4- [1- (4-cyclohexylphenyl) -3- (4-trifluoromethylsulfanylphenyl) ureidomethyl] benzoic acid methyl ester

4-((4-cyclohexylphenylamino) methyl) benzoic acid methyl ester (0.32 g, 1 mmol) is suspended in acetonitrile (5 mL) and 4- (trifluoromethylthio) phenyl isocyanate (0.24 g, 1.1 mmol) ) Was added. An additional amount of isocyanate (0.05 g) was added after day 1 and again (0.05 g) after day 2. On day 3 the reaction was stopped and concentrated in vacuo. Purify the residue by column chromatography on silica gel (30 g) using ethyl acetate: n-heptane (400 mL 1: 4 and 100 mL 1: 1) as eluent to afford 0.53 g of 4- [1- (4-cyclo Hexylphenyl) -3- (4-trifluoromethylsulfanylphenyl) ureidomethyl] benzoic acid methyl ester was obtained.

¹ H-NMR (DMSO- d ₆ ): δ 1.16-1.43 (m, 5H), 1.65-1.82 (m, 5H), 3.84 (s, 3H), 4.99 (s, 2H), 8.62 (s, 1 H); HPLC-MS (Method B): m / z = 543 (M + l); R _t = 9. 35 min.

Step 2a: 4-[-1- (4-cyclohexylphenyl) -3- (4-trifluoromethylsulfanylphenyl) ureidomethyl] benzoic acid

4- [1- (4-cyclohexylphenyl) -3- (4-trifluoromethylsulfanylphenyl) ureidomethyl] benzoic acid methyl ester (0.53 g, 0.98 mmol) in ethanol (96%, 11 mL) Dissolve and add sodium hydroxide (4 N, 1.47 mL). The mixture was stirred overnight. The reaction was dried over concentrated, water (15 mL) was added, acidified to pH 2-3 with hydrochloric acid (4 N, 1.6 mL) and extracted with ethyl acetate (25 mL). The aqueous phase was extracted once more with ethyl acetate (15 mL) and the combined organic phases were washed three times with water (10 mL), dried over magnesium sulfate, filtered and concentrated in vacuo. Crystals from ethyl acetate: n-heptane yielded 0.34 g of 4- [1- (4-cyclohexylphenyl) -3- (4-trifluoromethylsulfanylphenyl) ureidomethyl] benzoic acid.

¹ H-NMR (DMSO- d ₆ ): δ 1.5-1.42 (m, 5H), 1.67-1.83 (m, 5H), 2.45 (m, 1H), 5.00 (s, 2H), 7.157.25 (dd , 4H), 7.40 (d, 2H), 7.54-7.63 (dd, 4H), 7.88 (d, 2H), 7.62 (s, 1H), 12.90 (broad, 1H), HPLC-MS (method B) m / z = 529 (M + l); R _t = 8.55 min; M. p. 162.0-164.0 ° C.

Theoretical analysis for C ₂₈ H ₂₇ F ₃ N ₂ 0 ₃ S:

C, 63.62%; H, 5.15%; N, 5.30%. Found:

C, 63.97%; H, 5.28%; N, 5.26%.

Step 3: (R) -3- {4- [1- (4-cyclohexylphenyl) -3- (4-trifluoromethylsulfanylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionate methyl ester

4- [1- (4-cyclohexylphenyl) -3- (4-trifluoromethylsulfanylphenyl) ureidomethyl] benzoic acid (0.32 g, 0.606 mmol) was dissolved in DMF (7 mL) and HOAt (0.10 g, 0.727 mmol) and EDAC (0.14 g, 0.727 mmol) were added. The mixture was stirred for 30 minutes. Then (R) -3-amino-2-hydroxypropionic acid methyl ester hydrochloride (0.14 g) and diisopropylethylamine (0.16 mL, 0.909 mmol) were added. The reaction was stirred overnight. The reaction mixture was transferred to a separatory funnel with ethyl acetate (30 mL) and water (15 mL) and extracted. The aqueous phase is extracted once more with ethyl acetate (15 mL) and the combined organic phases are washed with hydrochloric acid (0.2 N, 3 x 10 mL) and an aqueous solution of saturated sodium chloride (3 x 10 mL) and then dried over magnesium sulfate, Filtered and concentrated in vacuo. 0.33 g of (R) -3- {4 was purified by purification of the residue on a column (silica gel, 30 g) using a mixture of ethyl acetate and n-heptane (200 mL, 40: 60 and 450 mL 1: 1) as eluent. -[1- (4-cyclohexylphenyl) -3- (4-trifluoromethylsulfanylphenyl) -ureidomethyl] benzoylamino} -2-hydroxypropionic acid methyl ester was obtained.

¹ H-NMR (DMSOd ₆ ) δ 1. 16-1.43 (m, 5H), 1.66-1.81 (m, 5H), 2.47 (m, 1H), 3.4 (m, 1H), 3.51 (m, 1H), 3.63 (s, 3H), 4.22 (q , 1 H), 4.97 (s, 2H), 5.73 (d, 1 H), 7.2 (dd, (4H), 7.35 (d, 2H), 7.60 (dd, 4H), 7.76 (d, 2H), 8.50 (t, 1 H), 8. 60 (s, 1 H), HPLC-MS ( method B): m / z = 630 (m + 1); R t = 8.07 min.

Step 4:

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (4-trifluoromethylsulfanylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid methyl ester (0.32 g, 0.508 mmol) was dissolved in ethanol (15 mL) and sodium hydroxide (4 N, 0.76 mL, 3.05 mmol) was added. The reaction mixture was stirred for 1.5 hours. The reaction was evaporated and water (15 mL) was added and then acidified with hydrochloric acid (4 N, 0.8 mL). The mixture was extracted with ethyl acetate (20 mL). The aqueous phase was extracted once more with ethyl acetate (15 mL) and the combined organic phases were washed with water (3 x 10 mL), dried over magnesium sulfate, filtered and concentrated to give the title compound (0.3 g).

¹ H-NMR (DMSO- d ₆ ): δ1.12-1.42 (m, 5H), 1.66-1.82 (m, 5H), 2.45 (m, 1H), 3.38 (m, 1H), 3.54 (m, 1H ), 4.17 (q, 1H), 4.96 (s, 2H), 5.45 (broad, 1H), 7.20 (dd, 4H), 7.34 (d, 2H), 7.60 (dd, 4H), 7.78 (d, 2H) , 8.45 (t, 1 H), 8.60 (s, 1 H), 12.53 (broad, 1 H), HPLC-MS (method B): m / z = 616 (M + l); R _t = 7.68 min.

Theoretical analysis for C ₃₁ H ₃₂ F ₃ N ₃ 0 ₅ S:

C, 60.48%; H, 5.24%; N, 6.83%. Found:

C, 60.25%; H, 5.52%; N, 6.53%.

Example 25 (General Method (A))

(R) -3- {4- [1- (4-cyclohexen-1-ylphenyl) -3- (3-methanesulfonyl-4-trifluoromethoxyphenyl) ureidomethyl] benzoylamino} -2 Hydroxypropionic acid

Preparation of Intermediate DN = C = O, 3-Methylsulfonyl-4-trifluoromethoxyphenyl Isocyanate Used in Step 2 :

To a solution of methyl iodine (59.0 g, 0.41 mol) in DMF (150 mL) was added potassium carbonate (23.0 g, 0.16 mol). 2- (trifluoromethoxy) thiophenol (16.0 g, 0.08 mol) was added in portions over 30 minutes. The reaction mixture was then stirred vigorously overnight. Water (250 mL) was added. The reaction mixture was extracted with ethyl acetate (2 x 150 mL). The combined organic phases were washed with 50% saturated aqueous sodium chloride solution (4 x 100 mL), dried (magnesium sulfate) and concentrated in vacuo to give 15.0 g of 1-methylsulfanyl-2-trifluoromethoxybenzene. .

Dissolve 1-methylsulfanyl-2-trifluoromethoxybenzene (15.0 g, 72 mmol) in DCM (200 mL) and add m-chloroperoxybenzoic acid (39.0 g, 216 mmol) in portions for 30 minutes. It was. The reaction mixture was then left overnight. DCM (200 mL) was added followed by the slow addition of sodium hydroxide (2 N, 200 mL). The organic phase was separated, washed with sodium hydroxide (2 N, 3 x 150 mL), dried (magnesium sulfate) and concentrated in vacuo to afford 15.8 g of 1-methylsulfonyl-2-trifluoromethoxybenzene.

^OneH-NMR (400 MHz, CDCl₃): δ8. 11 (d, 1H), 7.71 (t, 1H), 7.48 (m, 2H) 3.23 (s 1H); M. p. 44-46 ° C.

Theoretical Analysis for C ₈ H ₇ F ₃ 0 ₃ S

C, 40.00%; H, 2.94%. Found:

C, 40.22%; H, 2.92%.

1-methylsulfonyl-2-trifluoromethoxybenzene (15.7 g, 65 mmol) was dissolved in concentrated sulfuric acid (27 mL) and the solution was heated to 40 ° C. Nitric acid (100%, 27 mL) was added dropwise over 45 minutes. The reaction mixture was left overnight at 60 ° C., cooled and poured into crushed ice (300 mL). The precipitated product was isolated by filtration, washed with water (10 x 50 mL) and dried (magnesium sulfate) to give 17.5 g of 3-methylsulfonyl-4-trifluoromethoxynitrobenzene.

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 8.69 (d, 1H), 8.64 (d, 1H), 7.95 (d, 1H) 3.45 (s 3H); M. p. 102-104 ° C.

Theoretical Analysis for C ₈ H ₆ F ₃ NO ₅ S

C, 33.69%; H, 2. 12%; N, 4.91%. Found:

C, 33.91%; H, 2.08%; N, 4.92%.

3-methylsulfonyl-4-trifluoromethoxynitrobenzene (17.5 g) was dissolved in methanol (400 mL) and then palladium on carbon (10%, 50% water, 3.2 g) was added. The reaction mixture was hydrogenated at 1 atmosphere of hydrogen for 17 hours, filtered and concentrated in vacuo to yield 14.3 g of 3-methylsulfonyl-4-trifluoromethoxyaniline.

¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 7.26 (d, 1H), 7.14 (d, 1H), 6.85 (dd, 1H) 5.89 (s, 2H) 3.21 (s, 3H); M. p. 106-109 ° C.

Theoretical Analysis for C ₈ H ₈ F ₃ NO ₃ S:

37.65% C, 3.16% H, 5.49% N. Found:

37.65% C, 3.14% H, 5.45% N.

To 3-methylsulfonyl-4-trifluoromethoxyaniline (2.0 mmol, 500 mg) dissolved in ethyl acetate (6 mL) was added 3N hydrochloric acid in ethyl acetate (5 mL) and then concentrated in vacuo. The residue was treated three times with toluene (5 mL) and concentrated in vacuo each time. Toluene (10 mL) and diphosgene (6 mmol, 0.73 mL) were added to the residue under nitrogen atmosphere and the suspension was gently refluxed for 2 hours. Additional diphosgene (6 mmol, 0.73 mL) was added and refluxing continued overnight. The reaction mixture was concentrated in vacuo to afford 3-methylsulfonyl-4-trifluoromethoxyphenyl isocyanate.

Step 3: (R) -3- {4- [1- (4-cyclohexen-1-ylphenyl) -3- (3-methanesulfonyl-4-trifluoromethoxyphenyl) ureidomethyl] benzoylamino } -2-hydroxypropionic acid methyl ester

This intermediate was prepared using the general method (A) (steps 1,2,2a, and 3).

¹ H-NMR (DMSO- d ₆ ): δ 1.60 (m, 2H), 1.72 (m, 2H), 2.18 (m, 2H), 2.36 (m, 2H), 3.27 (s, 3H), 3.41 ( m, 1H), 3.51 (m, 1H), 3.61 (s, 3H), 4.23 (q, 1H), 4.96 (s, 2H), 5.70 (d, 1H), 6.18 (m, 1H), 7.19 (d , 2H), 7.33 (d, 2H), 7.39 (d, 2H), 7.53 (d, 1H), 7.75 (d, 2H), 8.00 (d, 1H), 8.18 (s, 1H), 8.50 (t , 1 H), 8.85 (s, 1 H); HPLC-MS (Method B): m / z = 690 (M + l); R _t = 6.92 min.

Step 4:

(R) -3- {4- [1- (4-cyclohexen-1-ylphenyl) -3- (3-methanesulfonyl-4-trifluoromethoxyphenyl) ureidomethyl] benzoylamino} -2 Hydrolysis of methyl hydroxypropionate gave the title compound .

¹ H-NMR (DMSO- d ₆ ): δ.60 (m, 2H), 1.72 (m, 2H), 2.18 (m, 2H), 2.36 (m, 2H), 3.27 (s, 3H), 3.41 ( m, 1H), 3.51 (m, 1H), 4.17 (t, 1H), 4.96 (s, 2H), 5.50 (broad, 1H), 6.18 (m, 1H), 7.19 (d, 2H), 7.33 (d , 2H), 7.39 (d, 2H), 7.53 (d, 1H), 7.75 (d, 2H), 8.00 (d, 1H), 8.18 (s, 1H), 8.50 (t, 1H), 8.85 (s , 1 H), 12.55 (broad, 1 H); HPLC-MS (Method B): m / z = 676 (M + l); R _t = 6.50 min.

Example 26 (General Method (A))

Trans- (R) -3- {4-[-3- (3,5-bis (methyl) phenyl) -1- (4-tert-butylcyclohexyl) ureidomethyl] benzoylamino} -2-hydroxy Propionic acid

Steps 1 and 2: trans-4- [3- (3,5-bis (methyl) phenyl) -1- (tert-butylcyclohexyl) ureidotyl] benzoic acid methyl ester

¹ H-NMR (DMSO- d ₆ ): δ0.82 (s, 9H), 0.93 (m, 1H), 1.11 (m, 2H), 1.41 (m, 2H), 1.71 (m, 4H), 2.23 ( s, 6H), 3.83 (s, 3H), 4.09 (m, 1H), 4.61 (s, 2H), 6.60 (s, 1H), 7.08 (s, 2H), 7.38 (d, 2H), 7.90 ( d, 2H), 8.20 (s, 1H); HPLC-MS (Method B): m / z = 451 (M + l); R _t = 8.93 min.

Step 2a: trans-4- [3- (3,5-bis (methyl) phenyl) -1- (tert-butylcyclohexyl) ureidomethyl] benzoic acid

Compounds were prepared by hydrolysis of trans-4- [3- (3,5-bis (methyl) phenyl) -1- (tert-butylcyclohexyl) ureidomethyl] benzoic acid methyl ester.

¹ H-NMR (DMSO- d ₆ ): δ0.82 (s, 9H), 0.93 (m, 1H), 1.11 (m, 2H), 1.41 (m, 2H), 1.71 (m, 4H), 2.23 ( s, 6H), 4.09 (m, 1H), 4.61 (s, 2H), 6.60 (s, 1H), 7.08 (s, 2H), 7.38 (d, 2H), 7.90 (d, 2H), 8.20 (s, 1 H), 12.82 (s, 1 H); HPLC-MS (Method B): m / z = 437 (M + l); R _t = 8.00 min.

Theoretical analysis for C ₂₇ H ₃₆ N ₂ 0 ₃ :

C, 74.28%; H, 8.31%; N, 6.42%. Found:

C, 74. 31%; H, 8.40%; N, 6.35%.

Step 3: trans- (R) -3- {4-[-3- (3,5-bis (methyl) phenyl) -1- (4-tert-butylcyclohexyl) ureidomethyl] benzoylamino} -2 Hydroxypropionic acid methyl ester:

Compounds were prepared from trans-4- [3- (3,5-bis (methyl) phenyl) -1- (tert-butylcyclohexyl) ureidomethyl] benzoic acid.

¹ H-NMR (DMSO- d ₆ ): δ0.82 (s, 9H), 0.93 (m, 1H), 1.11 (m, 2H), 1.43 (m, 2H), 1.71 (m, 4H), 2.23 ( s, 6H), 3.42 (m, 1H), 3.52 (m, 1H), 3.63 (s, 3H), 4.05 (m, 1H), 4.23 (q, 1H), 4.59 (s, 2H) , 5.70 (d, 1H), 6.58 (s, 1H), 7.08 (s, 2H), 7.30 (d, 2H), 7.78 (d, 2H), 8.18 (s, 1H), 8. 47 (t, 1H ); HPLC-MS (Method B): m / z = 538 (M + l); R _t = 7.43 min; M. p. 159-160 ° C.

Microanalytical Theory for C ₃₁ H ₄₃ N ₃ O ₅ :

C, 69.25%; H, 8.06%; N, 7. 81%. Found:

C, 69.03%; H, 8. 15%; N, 7.79%.

Step 4:

Trans- (R) -3- {4-[-3- (3,5-bis (methyl) phenyl) -1- (4-tert-butylcyclohexyl) ureidomethyl] benzoylamino} -2-hydroxy Hydrolysis of propionic acid methyl ester afforded the title compound.

¹ H-NMR (DMSO- d ₆ ): δ0.82 (s, 9H), 0.93 (m, 1H), 1.11 (m, 2H), 1.43 (m, 2H), 1.71 (m, 4H), 2.23 ( s, 6H), 3.40 (m, 1H), 3.55 (m, 1H), 4.05 (m, 1H), 4.18 (t, 1H), 4.59 (s, 2H), 6.58 (s, 1H), 7.08 (s , 2H), 7.30 (d, 2H), 7.78 (d, 2H), 8. 18 (s, 1H), 8.47 (t, 1H); HPLC-MS (Method B): m / z = 524 (M + l); R _t = 7.08 min.

Theoretical analysis for C ₃ 0H ₄₁ N ₃ 0 ₅ , 1½H ₂ O:

C, 65.43%; H, 8.05%; N, 7.63%. Found:

C, 65.54%; H, 7.93%; N, 7.44%.

Example 27 (General Method (A))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

¹ H-NMR (CDCl ₃ ): δ7.64 (d, 2H), 7.49 (br s, 1H), 7.25-7.15 (m, 6H), 7.03 (d, 2H), 6.90 (m, 1H), 6.40 (s, 1 H), 4.75 (s, 2H), 4.30 (br s, 1 H), 3.80-3.60 (m, 2H), 2.49 (m, 1 H), 1.90-1.65 (m, 5H), 1.45-1.25 (m, 5 H); HPLC-MS (Method B): m / z = 584 (M + l); R _t = 5.28 min.

Example 28 (General Method (A))

(R)-(3- {4- [1- (4-cyclohex-1-enylphenyl) -3- (3-fluoro-5-trifluoromethylphenyl) ureidomethyl] benzoylamino} -2- Hydroxypropionic acid

¹ H-NMR (DMSO- d ₆ ): δ 8.75 (s, 1H), 8.42 (t, 1H), 7.75 (d, 4H), 7.45-7.30 (m, 4H), 7.20 (d, 3H), 6.20 (s, 1H), 4.96 (s, 2H), 4.15 (dd, 1H), 3.55 (m, 1H), 3.40 (m, 1H), 2.35 (brs, 2H), 2.15 (br s, 2H). 1.75-1.55 (m, 4H); HPLC-MS (Method B): m / z = 600 (M + l); R _t = 5.01 min.

Example 29 (General Method (A))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (3-methylsulfanylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

_{^{1 H-NMR (DMSO- d 6}} ): δ12. 2 (brs, 1H), 8.44 (t, 1H), 8.20 (s, 1H), 7.78 (m, 2H), 7.40 (s, 1H), 7.33 (m, 2H), 7.25-7.10 (m, 6H) , 6.84 (d, 1H), 4.95 (s, 1H), 4.15 (dd, 1H), 3.55 (m, 1H), 3.38 (m, 1H). 2.42 (s, 3 H) 1.85-1.65 (m, 5 H), 1.40-1.15 (m, 5 H); HPLC-MS (Method B): m / z = 562 (M + l); R _t = 4. 77 min.

Example 30 (General Method (A))

(R) -3- {4- [1- (4-cyclohex-1-enylphenyl) -3- (2,2,4,5-tetrafluoro-4 H -Benzo [1,3] dioxin-6-yl) ureidomethyl] benzoylamino-2-hydroxypropionic acid

¹ H-NMR (CDCl ₃ ): δ7.70-7.50 (m, 3H), 7.45-7.30 (m, 4H), 7.25-6.85 (m, 6H), 6.12 (s, 1H), 4.80 (s, 2H ), 4.28 (m, 1H), 3.70 (m, 2H), 2.40-2.00 (m, 4H), 1.70-1.55 (m, 4H); HPLC-MS (Method B): m / z = 644 (M + l); R _t = 5.13 min.

Example 31 (General Method (A))

3- {4- [1- (4-Cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2 (R) methoxypropionic acid:

Step 3:

4- [1- (4-Cyclohex-1-enylphenyl) -3- (3,5-dichlorophenyl) ureidomethyl] benzoic acid (500 mg, 1.0 mmol), HOBt (184 mg, 1.2 mmol) and EDAC (232 mg; 1.2 mmol) was dissolved in a mixture of DCM (4.0 mL) and DMF (1.0 mL). The clear solution was stirred at ambient temperature for 1 hour. Diisopropylethylamine (515 μL) after addition of a solution of 3-amino-2 (R) -methoxypropionic acid methyl ester hydrochloride (257 mg, 1.5 mmol) in DCM (2.0 mL) and DMF (0.2 mL) Was added. The mixture was stirred overnight at room temperature, then diluted with DCM (40 mL) and washed once with a mixture of labeled sodium chloride / water (1: 2). The organic phase was dried over anhydrous sodium sulfate and dried in vacuo.

Step 4:

The oil was dissolved in a mixture of THF (4.0 mL) and methanol (4.0 mL). 4 N aqueous sodium hydroxide (625 μL, 2.5 mmol) was added and the mixture was stirred at rt for 2 h. The pH was adjusted to 3.0 with 1 N hydrochloric acid, and then the solvent was removed. The product was redissolved in ethyl acetate (20 mL) and washed once with water (20 mL). The aqueous phase was back-extracted with ethyl acetate (10 mL) and the combined organic phases were washed with sodium chloride (2 x 20 mL) and dried over anhydrous sodium sulfate. After removal of the solvent 230 mg (67%) of pure title compound were obtained.

_{^{1 H-NMR (DMSO- d 6}} ): δ12. 90 (bs, 1H), 8.55 (s, 1H), 8.54 (t, 1H), 7.76 (d, 2H), 7.63 (s, 2H), 7.41 (d, 2H), 7.34 (d, 2H), 7.20 (d, 2H), 7.15 (s, 1H), 6.18 (s, 1H), 4.95 (s, 2H), 3.90 (dd, 1H), 3.57 (m, 1H), 3.42 (m, 1H), 3.29 ( s, 3H), 2.34 (m, 2H), 2.16 (m, 2H), 1.70 (m, 2H), 1.59 (m, 2H); HPLC-MS (Method B): m / z = 596.2 (M + l); R _t = 5. 93 min.

Example 32 (General Method (A))

3- (4- {3- (3,5-dichlorophenyl) -1- [4- (2-methylcyclohex-1-enyl) phenyl] ureidomethyl} benzoylamino) -2- (R) -hydrate Roxypropionic acid and (R, S) -3- (4- {3- (3,5-dichlorophenyl) -1- [4- (6-methylchlorohex-1-enyl) phenyl] ureidomethyl} benzoylamino ) -2- (R) -hydroxypropionic acid

4- (2-methylcyclohex-1-enyl) aniline and (R, S) -4- (6-methylcyclohex-1-enyl) aniline (building block 11) and (R according to general method (A) The title compound was obtained using a mixture of) -3-amino-2-hydroxypropionic acid methyl ester (building block 5).

(R, S) -3- (4- {3- (3,5-dichlorophenyl) -1- [4- (6-methylcyclohex-1-enyl) phenyl] ureidomethyl} benzoylamino) -2 (R) -hydroxypropionic acid:

¹ H-NMR (DMSO- d ₆ ): δ 1,55 (s, 3H), 1,63 (bs, 4H), 2,03 (bs, 2H), 2,19 (bs, 2H), 3,47 (dm, 2H), 4,16 (m, 1H), 4,96 (s, 2H), 5,49 (bs, 1H), 7,15 (m, 5H), 7,33 (d, 2H) , 7,61 (s, 2H), 7,78 (d, 2H), 8,45 (t, 1H), 8,65 (s, 1H), 12,53 (bs, 1H); M. p. : 105-107 C; HPLC-MS (Method B): m / z = 596 (M < + >); Rt = 5,34 min.

3- (4- {3- (3,5-dichlorophenyl) -1- [4- (6-methylcyclohex-1-enyl) phenyl] ureidomethyl} benzoylamino) -2 (R) -hydroxy Propionic Acid:

¹ H-NMR (DMSO- d ₆ ): δ 0,90 (ds, 3H), 1,63 (bs, 4H), 2,03 (bs, 2H), 2,19 (bs, 2H), 3,47 (dm, 2H), 4,16 (m, 1H), 4,96 (s, 2H), 5,49 (bs, 1H), 5,93 (t, 1H), 7,15 (m, 5H) , 7,33 (d, 2H), 7,61 (s, 2H), 7,78 (d, 2H), 8,45 (t, 1H), 8,62 (s, 1H), 12,53 ( bs, 1 H).

Example 33 (General Method (A))

3- {4- [1- [4- (4-tert-butylcyclohex-1-enyl) phenyl] -3- (3,5-dichlorophenyl) ureidomethyl] benzoylamino} -2- (R) Hydroxypropionic acid

According to the general method (A), 4- (4-tert-butylcyclohex-1-enyl) aniline (building block 12) and (R) -3-amino-2-hydroxypropionic acid methyl ester (building block 5) To obtain the title compound .

¹ H-NMR (DMSO- d ₆ ): δ 0,88 (s, 9H), 1,23 (m, 2H), 1,93 (m, 2H), 2,27 (m, 3H), 3,47 (dm, 2H), 4,16 (dd, 1H), 4,95 (s, 2H), 6,19 (m, 1H), 7,13 (m1H), 7,18 (d, 2H), 7 , 33 (d, 2H), 7,39 (d, 2H), 7,62 (s, 2H), 7,77 (d, 2H), 8, 44 (t, 1H), 8,55 (s, 1H), 12, 53 (bs, 1H); M. p .: 151-155 C; HPLC-MS (Method B): m / z = 638 (M < + >); R _t = 6,04 min.

Example 34 (General Method (A))

(R, S) -3- (4- (3- (3,5-dichlorophenyl) -1- (4- (5-methylcyclohex-1-enyl) phenyl) ureidomethyl) -benzoylamino)- 2-hydroxypropionic acid and (R, S) -3- (4- (3- (3,5-dichlorophenyl) -1- (4- (3-methylcyclohex-1-enyl) phenyl) ureidomethyl ) Benzoylamino) -2-hydroxypropionic acid

(R, S) -4- (5-methylcyclohex-1-enyl) aniline and (R, S) -4- (3-methylcyclohex-1-enyl) aniline (building according to general method (A) Block 13) was used to obtain a mixture of the title compound (6: 4).

(R, S) -3- (4- (3- (3,5-dichlorophenyl) -1- (4- (5-methylcyclohex-1-enyl) phenyl) ureidomethyl) benzoylamino) -2 Hydroxypropionic acid:

¹ H-NMR (200 MHz, DMSO- d ₆ ): δ1.02 (d, 3H), 1.15-1-24 (m, 1H), 1.61-1.96 (m, 3H), 2.142.44 (m, 3H), 3.40 (t, 2H), 3.47-3.62 (m, 1H), 4.10-4.19 (m, 1H), 4.95 (s, 2H), 6.16 (t, 1H), 7.14 (t, 1H), 7.17 (d, 2H), 7.34 (d, 2H), 7.39 (d, 2H), 7.61 (d, 2H), 7.76 (d, 2H), 8.44 (t, 1H), 8.56 (s, 1H), 12.08 ( s br, 1H).

(R, S) -3- (4- (3- (3,5-dichlorophenyl) -1- (4- (3-methylcyclohex-1-enyl) phenyl) ureidomethyl) benzoylamino) -2 Hydroxypropionic acid:

¹ H-NMR (200 MHz, DMSO- d ₆ ): δ1.02 (d, 3H), 1.15-1-24 (m, 1H), 1.61-1.96 (m, 3H), 2.142.44 (m, 3H) , 3.40 (t, 2H), 3.47-3.62 (m, 1H), 4.10-4.19 (m, 1H), 4.95 (s, 2H), 6.04 (d, 1H), 7.14 (t, 1H), 7.17 (d , 2H), 7.34 (d, 2H), 7.39 (d, 2H), 7.61 (d, 2H), 7.76 (d, 2H), 8.44 (t, 1H), 8.53 (s, 1H), 12.08 (s br , 1H).

Example 35

3- {4- [3- [1 (S)-(4-Chlorophenyl) ethyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2 (R) -hydroxypropionic acid

4-[(4-cyclohexylphenylamino) methyl] benzoic acid methyl ester

Methyl 4-formylbenzoate (47 g, 285 mmol) was dissolved in methanol (400 mL) and 4-cyclohexylaniline (50 g, 0.285 mmol) in methanol (200 mL) was added slowly with mechanical stirring. More methanol (1 L) was added and the suspension was stirred for 3 days at room temperature. Filtration, washing and drying in vacuo gave 90.7 g (99%) of 4 [(4-cyclohexylphenylimino) methyl] benzoic acid methyl ester. It was dissolved in N-methylpyrrolidine (855 mL) and methanol (45 mL). The temperature was kept below 40 ° C. with mechanical stirring and sodium borohydride pellets (42.4 g, 1.12 mol) were added in portions. The mixture was stirred for 2 h at room temperature and 16 h at 40 ° C. The mixture was cooled to 5 ° C. and water (2 L) was added slowly. Acetone (350 mL) was then added and the mixture was stirred at 5 ° C. for 1 hour. Filtration, washing with water (2 x 500 mL) and drying in vacuo gave 78 g (86%) of 4-[(4-cyclohexylphenylamino) methyl] benzoic acid methyl ester in solid form.

¹ H-NMR (CDCl ₃ ): δ 1.2-1.4 (5H, m), 1.7-1.85 (5H, m), 2.39 (1H, m), 3.97 (3H, s), 4.04 (1H, bs), 4.39 (2H, s), 6.55 (2H, d), 7.01 (2H, d), 7.44 (2H, d), 8.00 (2H, d).

N-chlorocarbamoyl-4-[(4-cyclohexylphenylamino) methyl] benzoic acid methyl ester

4-[(4-cyclohexylphenylamino) methyl] benzoic acid methyl ester (75 g, 0.23 mol) was dissolved in THF (750 mL). Diisopropylethylamine (56.0 mL, 0.32 mmol) and 4-dimethylaminopyridine (1.0 g; 8.1 mmol) were added. The solution was cooled to 5 ° C. Bis (trichloromethyl) carbonate (28.0 g, 0.093 mol) was added portionwise while maintaining the internal reaction temperature below 10 ° C. The mixture was further stirred at 10 ° C. for 2 h and then transferred to a separatory funnel. Ethyl acetate (800 mL) and water (1000 mL) were added. After mixing, the organic layer was separated, dried over anhydrous sodium sulfate, dried by vacuum evaporation and concentrated. The product was obtained quantitatively as a stable, hard crystalline material.

¹ H-NMR (CDCl ₃ ): δ7.92 (d, 2H); 7.40 (d, 2 H); 7.25 (d, 2 H); 7.17 (d, 2 H); 4.98 (s, 2 H); 3.83 (s, 3 H); 2.5 (m, 1 H); 1.65-1.80 (m, 5H); 1.15-1.40 (m, 5H).

N-chlorocarbamoyl-4-[(4-cyclohexylphenylamino) methyl] benzoic acid methyl ester

4-[(4-cyclohexylphenylamino) methyl] benzoic acid methyl ester (75 g, 0.23 mol) was dissolved in THF (750 mL). Diisopropylethylamine (56.0 mL, 0.32 mmol) and 4-dimethylaminopyridine (1.0 g, 8.1 mmol) were added. The solution was cooled to 5 ° C. Bis (trichloromethyl) carbonate (28.0 g, 0.093 mol) was added portionwise while maintaining the internal reaction temperature below 10 ° C. The mixture was further stirred at 10 ° C. for 2 h and transferred to a separatory funnel. Acetyl acetate (800 mL) and water (1000 mL) were added. After mixing, the organic layer was separated, dried over anhydrous sodium sulfate, dried by rotary evaporation in vacuo and concentrated. The product was obtained as a stable, hard crystalline material.

¹ H-NMR (CDCl ₃ ): δ7.92 (d, 2H), 7.40 (d, 2H), 7.25 (d, 2H), 7.17 (d, 2H), 4.98 (s, 2H), 3.83 (s, 3H), 2.5 (m, 1H), 1.65-1.80 (m, 5H), 1.15-1.40 (m, 5H).

4- [3- [1 (S)-(4-chlorophenyl) ethyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid methyl ester

N-chlorocarbamoyl-4-[(4-cyclohexylphenylamino) methyl] benzoic acid methyl ester (94 g, 0.244 mol) and N-methyl-2-pyrrolidinone in a 2 L reaction flask equipped with mechanical stirring (1.0 L) and triethylamine (68 mL, 0.487 mol) were added. While maintaining the internal reaction temperature below 30 ° C., (S) -1- (4-chlorophenyl) ethylamine (38.0 g, 0.244 mol) was added dropwise to the clear solution, and the reaction mixture was then water (1.0 L) and Partitioned in ethyl acetate (1.0 L). After extensive mixing, the organic layer was separated and washed with 5% aqueous citric acid solution (500 mL) and with saturated ammonium chloride (500 mL) before drying over anhydrous sodium sulfate. Solvent was removed and residual oil was evaporated once from acetonitrile. The product was pure enough for future synthesis. Yield: 103 g (84%).

¹ H-NMR (DMSO- d ₆ ): δ 7.88 (d, 2H), 7.32 (d, 2H), 7.30 (d, 4H), 7.19 (d, 2H), 7.08 (d, 2H), 6.28 ( d, 1H), 4.88 (dd, 2H), 4.76 (m, 1H), 3.81 (s, 3H), 2.44 (m, 1H), 1.65-1.80 (m, 5H), 1.151.40 (m, 5 H); HPLC-MS (Method B): m / z = 505 (M + l); Rt = 6.17 min.

4- [3- [1 (S)-(4-Chlorophenyl) ethyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid

4- [3- [1 (S)-(4-chlorophenyl) ethyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid methyl ester (35.0 g, 69.3 mmol) in ethanol (400 mL) Dissolved. 4 N aqueous sodium hydroxide (100 mL) was added and the clear solution was stirred at rt for 3 h. The solution was neutralized with 4 N hydrochloric acid (100 mL) and placed on an ice bath to start crystallization. The crystals were collected, washed extensively with water and then dried in vacuo overnight. Yield 34.25 g (100%).

¹ H-NMR (DMSO- d ₆ ): δ 12.85 (bs, 1H), 7.85 (d, 2H), 7.32 (d, 2H), 7.30 (d, 4H), 7.19 (d, 2H), 7.08 ( d, 2H), 6.27 (d, 1H), 4.85 (m, 3H), 2.45 (m, 1H), 1.65-1.80 (m, 5H), 1.15-1.40 (m, 5H); HPLC-MS (Method B): m / z = 491 (M + l); Rt = 5.50 min.

3- {4- [3- [1 (S)-(4-chlorophenyl) ethyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2 (R) -hydroxypropionic acid methyl ester

4- [3- [1 (S)-(4-chlorophenyl) ethyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoic acid (200 mg, 0.4 mmol), HOBt (75 mg, 0.5 mmol) , And EDAC (94 mg, 0.5 mmol) were dissolved in a mixture of DMF (200 L) and DCM (2 mL). The clear solution was stirred at room temperature for 90 minutes. A solution of R-isoserine methyl ester hydrochloride (95 mg, 0.6 mmol) in DCM (1.0 mL) and DMF (0.4 mL) was added and the reaction mixture was stirred overnight at ambient temperature. The reaction mixture was partitioned between DCM (20 mL) and water (20 mL). The organic phase was separated and washed with a mixture of brine and water (1: 2), then dried over anhydrous sodium sulfate and evaporated to dryness. The residue was evaporated sequentially from acetonitrile to give quantitative yield of the title material.

¹ H-NMR (DMSO- d ₆ ): δ 8.48 (t, 1H), 7.73 (d, 2H), 7.34 (d, 2H), 7.30 (d, 2H), 7.24 (d, 2H), 7.18 ( d, 2H), 7.08 (d, 2H), 6.27 (d, 1H), 5.70 (d, 1H), 4.34 (m, 1H), 4.32 (d, 2H), 4.22 (q, 1H), 3.62 (s, 3H), 3.52 (m, 1H), 3.40 (m, 1H), 1.65-1.80 (m, 5H), 1.10-1.40 (m, 9H).

3- {4- [3- [1 (S)-(4-Chlorophenyl) ethyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2 (R) -hydroxypropionic acid

3- {4- [3- [l (S)-(4-chlorophenyl) ethyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2 (R) -hydroxypropionic acid methyl ester (280 mg, 0.473 mmol) was dissolved in a mixture of THF (2.5 mL) and methanol (2.5 mL) and 4N aqueous sodium hydroxide (0.355 mL) was added. The reaction mixture was stirred at rt for 2 h. The pH was adjusted to 3.0 with 1 N hydrochloric acid. The solvent was removed by rotary evaporation in vacuo and the residue was redissolved in ethyl acetate (10 mL). The organic phase was washed twice with water and once with brine, then concentrated in vacuo to dryness to afford the title compound as a powder. Yield: 168 mg (89%).

¹ H-NMR (DMSO- d ₆ ): δ 8.46 (t, 1H), 7.75 (d, 2H), 7.35 (d, 2H), 7.31 (d, 2H), 7.25 (d, 2H), 7.19 ( d, 2H), 7.08 (d, 2H), 6.28 (d, 2H), 4.85 (m, 1H), 4.80 (d, 2H), 4.15 (m, 1H), 3.55 (m, 1H), 3.40 (m , 1H), 1.65-1.80 (m, 5H), 1.10-1.40 (m, 9H); HPLC-MS (Method B): m / z = 579 (M + l); Rt = 5.27 min.

Example 36

3- {4- [3-biphenyl-2-ylmethyl-1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2 (R) hydroxypropionic acid

The compound is similar to that described in Example 35, with N-chlorocarbamoyl-4-[(4-cyclo) prior to hydrolysis of benzoic acid methyl ester, which is coupled with (R) -isoserine ethyl ester hydrochloride. Hexylphenylamino) methyl] benzoic acid methyl ester and biphenyl-2-ylmethylamine. Hydrolysis gave the title compound .

_{^{1 H-NMR (DMSO- d 6}} ): δ12. 6 (s, 1H), 8.45 (t, 1H), 7.78 (d, 2H), 7.45-7.20 (m, 14H), 7.05 (d, 2H), 6.10 (t, 1H), 5.50 (bs, 1H) , 4.85 (s, 2H), 4.2 (m, 3H), 3.55 (m, 1H), 2.45 (m, 1H), 1.851.70 (m, 5H), 1.40-1.20 (m, 6H); HPLC-MS (Method B): m / z = 606 (M + l); Rt = 5.08 min.

General method (B) for solution phase synthesis of compounds of formulas (Ia) and (Ib):

Wherein R ² , R ³ , R ⁷ , R ⁸ , A, E and D are as defined in formula (I).

Step 6 is performed using 1) BSA and 2) D-N = C═O when A is —CHOH—. Otherwise, step 6 is performed using only D-N = C = O.

The method is further described in the following examples.

Example 37 (General Method (B))

(R) -3- {4- [3- (4-cyano-3-trifluoromethylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

Step 1 is carried out using the same method as in general method (A).

Step 2: 4-{[tert-butoxycarbonyl- (4-cyclohexylphenyl) amino] methyl} benzoic acid methyl ester

4-((4-cyclohexylphenylamino) methyl) benzoic acid methyl ester (2.0 g, 6.18 mmol) is suspended in sodium hydroxide (1 N, 6.18 mL) and di-tert-butyldicarbonate in THF (10 mL) ( 1.67 g, 7.42 mmol) was added dropwise. The reaction mixture was stirred overnight and concentrated in vacuo to a solid residue, which was redissolved in diethyl ether (50 mL) and washed with water (25 mL) added with sodium hydroxide (1.3 mL, 1 N). . The aqueous phase was extracted again with diethyl ether (25 mL) at pH 11-12. The combined organic phase was washed with sodium hydrogen sulfate (30 mL, 10%) and water (3 x 20 mL), then dried over magnesium sulfate and concentrated in vacuo. Crystal formation from ethyl acetate and n-heptane yielded 1.98 g of 4-{[tert-butoxycarbonyl- (4-cyclohexylphenyl) amino] methyl} benzoic acid methyl ester.

¹ H-NMR (DMSO- d ₆ ): δ1.13-1.44 (m, 14H), 1.63-1.81 (m, 5H), 2.46 (m, 1H), 3.83 (s, 3H), 4.88 (s, 2H ), 7.12 (m, 4H), 7. 48 (d, 2H), 7.92 (d, 2H); HPLC-MS (Method B): m / z = 424 (M + l); R _t = 9.10 min; M. p. 99.5-101.0 ° C.

Theoretical analysis for C ₂₆ H ₃₃ NO ₄ :

C, 73.73%; H, 7.85%; N, 3.31%. Found:

C, 73.30%; H, 8.07%; N, 3.26%.

Step 3: 4-{[tert-butoxycarbonyl- (4-cyclohexylphenyl) amino] methyl} benzoic acid

4-{[tert-butoxycarbonyl- (4-cyclohexylphenyl) amino] methyl} benzoic acid methyl ester was suspended in ethanol (30 mL) and sodium hydroxide (4 N, 8.1 mL) was added. The reaction mixture was stirred overnight. The mixture was concentrated to dryness, suspended in water (100 mL), acidified with hydrochloric acid (8.5 mL, 4 N) and extracted with ethyl acetate (100 mL). The aqueous phase was extracted once more with ethyl acetate (30 mL) and the combined organic phases were washed with water (3 x 50 mL), then dried over magnesium sulfate and concentrated in vacuo. The residue was crystallized from a mixture of ethyl acetate and n-heptyl to yield 1.75 g of 4-{[tert-butoxycarbonyl- (4-cyclohexylphenyl) amino] methyl} -benzoic acid.

¹ H-NMR (CDCl ₃ -d ₆ ): δ 1. 18-1.42 (m, 14H), 1.68-1.87 (m, 5H), 2.46 (m, 1H), 4.88 (s, 2H), 7.10 (m, 4H), 7.47 (d, 2H), 8. 07 ( d, 2H); HPLC-MS (Method B): m / z = 410 (M + l); R _t = 8.15 min; M. p. 192.5-194.5 ° C.

Theoretical analysis for C ₂₅ H ₃₁ NO ₄ :

C, 73.32%; H, 7.63%; N, 3.42%. Found:

C, 73.03%; H, 7.86%; N, 3.36%.

Step 4: (R) -3- (4-{[tert-butoxycarbonyl- (4-cyclohexylphenyl) amino] methyl} benzoylamino) -2-hydroxypropionic acid methyl ester

4-{[tert-butoxycarbonyl- (4-cyclohexylphenyl) amino] methyl} benzoic acid is dissolved in DMF (10 mL) and HOBt (0.40 g, 2.93 mmol) and EDAC (0.52 g, 2.73 mmol) Added. The reaction mixture was stirred for 45 minutes. Then a solution of (R) -3-amino-2-hydropropionic acid methyl ester in DMF (8 mL) and diisopropylethylamine (0.46 mL) was added. The mixture was stirred overnight. The reaction mixture was diluted with water (40 mL) and extracted with ethyl acetate (75 mL). The aqueous phase was extracted with ethyl acetate (30 mL). The combined organic phase was washed with hydrochloric acid (0.2 N, 3 × 30 mL), water: saturated sodium chloride (3 × 30 mL), dried over magnesium sulfate and concentrated in vacuo. 0.77 g of (R)-was purified by column chromatography on silica gel (100 g) using ethyl acetate and n-heptane (1 L (1: 1) and 0.5 L (7: 3)) as eluent. 3- (4-{[tert-butoxycarbonyl- (4-cyclohexylphenyl) amino] methyl} benzoylamino) -2-hydropropionic acid methyl ester was obtained.

¹ H-NMR (DMSO- d ₆ ): δ 1.16-1.41 (m, 14H), 1.63-1.81 (m, 5H), 2.46 (m, 1H), 3.42 (m, 1H), 3.54 (m, 1H ), 3.62 (s, 3H), 4.24 (m, 1H), 4.84 (s, 2H), 5.70 (d, 1H), 7.12 (m, 4H), 7.28 (d, 2H), 7.78 (d, 2H) , 8.51 (t, 1 H); HPLC-MS (Method B): m / z = 511 (M + l); R _t = 7.63 min.

Step 5: (R) -3- {4-[(4-cyclohexylphenylamino) methyl] benzoylamino} -2-hydroxypropionic acid methyl ester

(R) -3- (4-{[tert-butoxycarbonyl- (4-cyclohexylphenyl) amino] methyl} benzoylamino) -2-hydroxypropionic acid methyl ester was dissolved in ethyl acetate (10 mL) Anhydrous hydrogen chloride in ethyl acetate (3 M, 10 mL) was added. The mixture was stirred at rt for 2 h and concentrated in vacuo. The residue was suspended in ethyl acetate (15 mL) and concentrated. This was repeated twice. The residue was then suspended in ethyl acetate (10 mL) and placed at 5 ° C. overnight. The precipitate was filtered off, washed with ice-cold ethyl acetate and dried in vacuo to give 0.62 g of (R) -3- {4-[(4-cyclohexylphenylamino) methyl] benzoylamino} -2-hydropropionic acid methyl ester Got.

¹ H-NMR (DMSO- d ₆ ): δ1.12-1.43 (m, 5H), 1.63-1.82 (m, 5H), 2.45 (m, 1H), 3.42 (m, 1H), 3.53 (m, 1H ), 3.60 (s, 3H), 4.25 (t, 1H), 4.48 (s, 2H), 7.18 (m, 4H), 7.57 (d, 2H), 7.82 (d, 2H), 8.58 (t, 1 H ); HPLC-MS (Method B): m / z = 411 (M + 1); R _t = 4. 93 min.

Step 6: (R) -3- {4- [3- (4-cyano-3-trifluoromethylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxy Propionic acid methyl ester

5-amino-2-cyanobenzotrifluoride (0.07 g, 0.36 mmol) was dissolved in ethyl acetate (2 mL) and anhydrous hydrogen chloride in ethyl acetate (3.5 M, 5.5 mL) was added. After 15 minutes the solution was concentrated to dryness and co-evaporated three times with toluene (3 × 5 mL). Toluene (2.5 mL) was added to the residue and flushed with nitrogen for about 10 minutes before adding diphosgene (0.43 mL). The mixture was then refluxed gently for 1 hour under a nitrogen atmosphere. The mixture was cooled, concentrated to dryness in vacuo and co-evaporated twice with toluene to remove excess diphosgene to afford 4-cyano-3-trifluoromethylphenyl isocyanate.

(R) -3- {4-[(4-cyclohexylphenylamino) methyl] benzoylamino} -2-hydropropionic acid methyl ester, hydrochloride (0.13 g, 0.3 mmol) is dissolved in DCM (5 mL) and BSA (0.22 mL, 0.9 mmol) was added. The mixture was stirred for 0.5 h and diisopropylethylamine (0.052 mL, 0.3 mmol) was added. The reaction mixture was added to the isocyanate and stirred overnight. The reaction mixture was transferred to a separatory funnel and washed twice with water (10 mL), then dried over magnesium sulfate and concentrated in vacuo. Purify the residue by column chromatography (30 g) using ethyl acetate / n-heptane (4: 6) (400 mL) and then ethyl acetate (200 mL) as eluent to afford 0.085 g of (R) -3- { 4- [3- (4-cyano-3-trifluoromethylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydropropionic acid methyl ester was obtained.

¹ H-NMR (DMSO- d ₆ ): δ1.12-1.44 (m, 6H), 1.66-1.82 (m, 5H), 3.41 (m, 1H), 3.53 (m, 1H), 3.60 (s, 3H ), 4.22 (m, 1H), 4.47 (s, 2H), 5.69 (s, 1H), 7.21 (m, 4H), 7.33 (d, 2H), 7.76 (d, 2H), 7.98 (s, 2H) , 8.14 (s, 1 H), 8.48 (t, 1 H), 9.1 (s, 1 H); HPLC-MS (Method B): m / z = 623 (M + l); R _t = 6.02 min.

Step 7:

(R) -3- {4- [3- (4-cyano-3-trifluoromethylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydropropionic acid methyl ester ( 0.07 g, 0.124 mmol) was suspended in ethanol (3 mL) and sodium hydroxide (4 N, 0.19 mL, 0.742 mmol) was added. The reaction mixture was stirred for 1.5 h and concentrated to remove ethanol. The residue was diluted with water (10 mL) and acidified with hydrochloric acid (4 N, 0.21 mL). The mixture was extracted with ethyl acetate (2 x 10 mL) and the combined organic phases were washed with water (3 x 10 mL), dried over magnesium sulfate and concentrated in vacuo to afford the title compound (0.68 g).

¹ H-NMR (DMSO- d ₆ ): δ 1.16-1.42 (m, 6H), 1.66-1.82 (m, 5H), 3.40 (m, 1H), 3.54 (m, 1H), 4.16 (m, 1H ), 4.48 (s, 2H), 7.20 (m, 4H), 7.34 (d, 2H), 7.78 (d, 2H), 7.99 (s, 2H), 8.16 (s, 1H), 8.44 (t, 1H) , 9.1 (s, 1 H); HPLC-MS (Method B): m / z = 609 (M + l); R _t = 7.27 min.

Example 38 (General Method (B))

(R) -3- {4- [3- (3-tert-butylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

Step 6: (R) -3- {4- [3- (3-tert-butylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino-2-hydropropionic acid methyl ester

3- (tert-butyl) aniline (0.054 g, 0.36 mmol) was dissolved in ethyl acetate (2 mL) and anhydrous hydrogen chloride in ethyl acetate was added twice (3.5 M, 3 mL + 2.5 mL). After 15 minutes the mixture was concentrated to dryness and co-evaporated three times with toluene (3 × 5 mL). Toluene (2.5 mL) was added to the residue and flushed with nitrogen for about 10 minutes before adding diphosgene (0.43 mL). The mixture was then refluxed gently for 1 hour under a nitrogen atmosphere. The mixture was cooled and concentrated in vacuo. This was repeated twice to remove excess dispogen. The mixture was concentrated to dryness and co-evaporated three times with toluene (5 mL each time). The residue was concentrated to dryness and co-evaporated twice from toluene. It was then redissolved in toluene (2.5 mL) and flushed with nitrogen for about 10 minutes before adding diphosgene (0.43 mL). The mixture was gently refluxed under nitrogen for 1 hour. After cooling, the mixture was concentrated and co-evaporated twice from toluene to remove excess diphosgene to afford 3-tert-butyl-phenyl isocyanate.

(R) -3- {4-[(4-cyclohexylphenylamino) methyl] benzoylamino} -2-hydropropionic acid methyl ester, hydrochloride (0.13 g, 0.3 mmol) is dissolved in DCM (5 mL) and BSA (0.22 mL, 0.9 mmol) was added. The mixture was stirred for 0.5 h and diisopropylethylamine (0.052 mL, 0.3 mmol) was added. The reaction mixture was added to the isocyanate and stirred overnight. The reaction was transferred to a separatory funnel, washed with water (10 mL), dried over magnesium sulfate and concentrated in vacuo. The residue was purified by column chromatography on silica gel (30 g) using ethyl acetate and n-heptane (6: 4) (400 mL) and then ethyl acetate (100 mL) as eluent to afford 0.12 g of (R)-. 3- {4- [3- (3-tert-butylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydropropionic acid methyl ester was obtained.

¹ H-NMR (DMSO- d ₆ ): δ1.23 (s, 11H), 1.28-1.42 (m, 3H), 1.65-1.80 (m, 5H), 2.47 (m, 1H), 3.40 (m, 1H ), 3.51 (m, 1H), 4.22 (m, 1H), 4.94 (s, 2H), 5.71 (d, 1H), 6.99 (d, 1H), 7.12-7-24 (m, 5H), 7.28 ( d, 1H), 7.36 (d, 2H), 7.42 (s, 1H), 7.77 (d, 2H), 8. 08 (s, 1H), 8.50 (t, 1H); HPLC-MS (Method B): m / z = (585 + 1); R _t = 8.30 min.

Step 7:

(R) -3- {4- [3- (3-tert-butylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydropropionic acid methyl ester (0.11 g, 0.188 mmol ) Was dissolved in ethanol (4 mL) and sodium hydroxide (4 N, 0.28 mL, 1.128 mmol) was added. The reaction was stirred for 1.5 hours and concentrated in vacuo to remove ethanol. The residue was diluted with water (10 mL), then acidified with hydrochloric acid (4 N, 0.3 mL) and extracted with ethyl acetate (2 x 10 mL). The combined organic phases were washed with water (3 × 10 mL), dried over magnesium sulfate and concentrated in vacuo to afford the title compound (0.10 g).

¹ H-NMR (DMSO- d ₆ ): δ1.23 (s, 9H), 1.28-1.42 (m, 4H), 1.65-1.81 (m, 5H), 2.47 (m, 1H), 3.38 (m, 1H ), 3.55 (m, 1H), 4.16 (m, 1H), 4.94 (s, 2H), 7.00 (d, 1H), 7.11-7.24 (m, 6H), 7.28 (d, 1H), 7.35 (d, 1 H), 7.41 (s, 1 H), 7.80 (d, 2 H), 8.10 (s, 1 H), 8.46 (t, 1 H); HPLC-MS (Method B): m / z = 572 (M + l); R _t = 7.78 min.

Example 39 (General Method (B))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (3-hydroxymethyl-4-trifluoromethoxyphenyl) ureidomethyl] benzoylamino} -2-hydropropionic acid

Preparation of 3- (tert-butyldimethylsilanyloxymethyl) -4-trifluoromethoxyaniline used in step 6:

Fuming nitric acid (5 mL) was cooled on an ice bath. Methyl 2- (trifluoromethoxy) benzoate (5 g, 22.7 mmol) was added slowly within 30 minutes while maintaining the temperature below 15 ° C. The reaction was stirred at 60 ° C. for 1 hour and at room temperature for 2 hours. The mixture was poured into ice water to separate the oil. The aqueous supernatant was drained off and additional water (50 mL) was added to the oil. After neutralization with sodium hydrogen carbonate, the mixture was extracted with ethyl acetate (25 mL). The aqueous phase was extracted once more with ethyl acetate (15 mL). The combined organic phase was washed with saturated sodium chloride (2 x 15 mL), then dried (magnesium sulfate) and concentrated in vacuo to give 5.69 g of 5-nitro-2-trifluoromethoxybenzoic acid methyl ester.

_{^{1 H-NMR (DMSO- d 6}} ): δ3.93 (3H, s), 7.82 (1H, d), 8.58 (1H, d), 8.67 (1H, s); HPLC-MS (Method B): m / z: 266; R _t = 6.0 min.

5-nitro-2-trifluoromethoxybenzoic acid methyl ester (5.69 g, 21.5 mmol) was dissolved in 99.9% (80 mL) of ethanol and tin (II) chloride dihydrate (24.2 g, 107 mmol) was added. The suspension was stirred for 2 h at 75 ° C. on an oil-bath and concentrated in vacuo. Ethyl acetate (100 mL) and water (50 mL) were added and the pH adjusted to 8 with 4N sodium hydroxide (50 mL). The liquid was poured from the precipitate. The precipitate was extracted twice with ethyl acetate. The aqueous phase was extracted twice with ethyl acetate (60 mL). The combined organic phases were washed with saturated sodium chloride solution (2 × 100 mL), dried (magnesium sulfate) and concentrated in vacuo. Purification by column chromatography (120 g silica) using ethyl acetate and heptane (1: 1) as an eluent gave 3.8 g of 5-amino-2-trifluoromethoxybenzoic acid methyl ester.

_{^{1 H-NMR (DMSO- d 6}} ): δ3.82 (3H, s), 5.63 (2H, s), 6.79 (1H, d), 7.07 (1H, s), 7.11 (1H, d); HPLC-MS (Method B): m / z: 236, R _t = 4.6 min.

5-amino-2-trifluoromethoxybenzoic acid methyl ester (3.0 g, 12.8 mmol) was dissolved under nitrogen in THF (20 mL) in a three-necked flask equipped with a thermometer and an additional funnel. Lithium aluminum hydride (1M in THF, 15 mL) was added dropwise within 10 minutes under stirring and ice cooling. Stirring was continued for 1 hour at room temperature and the reaction was concentrated in vacuo. The residue is suspended in DCM (150 mL) and water (50 mL), then filtered through celite and washed with DCM and water. The filtrate was removed and the aqueous phase was extracted once more with DCM (30 mL). The combined organic phase was washed with water (2 × 20 mL), then dried (magnesium sulfate) and concentrated in vacuo to afford 2.47 g of (5-amino-2-trifluoromethoxyphenyl) methanol.

_{^{1 H-NMR (DMSO- d 6}} ): δ3.92 (2H, d), 5.18 (1H, t), 5.28 (2H, s), 6.45 (1H, d), 6.91 (1H, d); HPLC-MS (Method B): m / z: 208, R _t = 7.2 min.

5-amino-2-trifluoromethoxyphenyl) methanol (1.2 g, 5.8 mmol) was dissolved in DMF (5 mL) and imidazole (0.48 g, 7.1 mmol) and tert-butyldimethylsilyl chloride (0.99 g, 6.6 mmol) was added. The reaction mixture was stirred for 16 h and water (20 mL) was added. The mixture is extracted with ethyl acetate (2 x 50 mL) and the combined organic phases are washed with water (10 mL), citric acid (10 mL, 10%) and water (2 x 10 mL), dried (magnesium sulfate) and vacuum Concentrated at. Purify the residue by column chromatography (110 g, silica) using ethyl acetate and heptane (1: 3) as eluent to obtain 1.2 g of 3- (tert-butyldimethylsilanyloxymethyl) -4-trifluorome The oxyaniline was obtained.

¹ H-NMR (DMSO- d ₆ ): δ0.82 (9H, s), 3.25 (6H, s), 4.52 (2H, s), 5.23 (2H, s), 6.41 (1H, d), 6.61 ( 1 H, s), 6.86 (1 H, d); HPLC-MS (Method B): m / z: 322; R _t = 7.17 min.

Step 6: (R) -3- {4- [3- [3- (tert-butyldimethylsilanyloxymethyl) -4-trifluoromethoxyphenyl] -1- (4-cyclohexylphenyl) ureidomethyl ] Benzoylamino} -2-hydroxypropionic acid methyl ester

Bis (trichloromethyl) carbonate (triphosgene) (0.09 g, 0.31 mmol) was dissolved in DCM (2 mL) and cooled in an ice bath under nitrogen. 3- (tert-butyldimethylsilanyloxymethyl) -4-trifluoromethoxyaniline (0.3 g, 0.93 mmol) was evaporated twice from toluene to remove all moisture and then dissolved in DCM (2 mL) and di Isopropylethylamine (0.32 mL) was added. Cooled tripphosgen was added to this solution and the mixture was stirred at 20 ° C. for 2.5 h. (R) -3- {4-[(4-cyclohexylphenylamino) methyl] benzoylamino} -2-hydroxypropionic acid hydrochloride (0.37 g, 0.83 mmol) was evaporated twice from toluene followed by DMF (3 mL) and diisopropylethylamine (0.141 mL, 0.83 mmol) were added. While stirring, the solution was added to the isocyanate and heated at 80 ° C. for 2 hours under nitrogen. The reaction mixture was evaporated in vacuo and the residue was extracted with DCM (80 mL), aqueous citric acid (10%, 25 mL). The aqueous phase was extracted with DCM (30 mL). The combined organic phase was washed with aqueous citric acid (10%, 3 x 25 mL), then dried over magnesium sulfate and evaporated in vacuo. Purify the residue by column chromatography on silica gel (30 g) using ethyl acetate and n-heptane (940 mL, 1: 1 and 300 mL ethyl acetate) as eluent to afford 0.03 g of (R) -3- {4 -[3- [3- (tert-butyldimethylsilanyloxymethyl) -4-trifluoromethoxyphenyl] -1- (4-cyclohexylphenylureidomethyl] benzoylamino} -2-hydropropionic acid methyl ester Got it.

HPLC-MS (Method B): m / z = 758 (M + l); R _t = 9.57 min.

Step 7:

(R) -3- {4- [3- [3- (tert-butyldimethylsilanyloxymethyl) -4-trifluoromethoxyphenyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino } -2-hydropropionic acid methyl ester (24 mg, 0.032 mmol) was dissolved in ethanol (1 mL) and sodium hydroxide (0.05 mL, 019 mmol) was added. The reaction mixture was stirred for 2 hours and concentrated to remove ethanol. The residue was diluted with water (10 mL), then acidified with hydrochloric acid (4 N, 0.3 mL) and extracted with ethyl acetate (2 x 10 mL). The combined organic phases were washed with water (3 x 10 mL), dried over magnesium sulfate and concentrated in vacuo to afford 17 mg of (R) -3- {4- [3- [3- (tert-butyldimethylsilanyloxymethyl ) -4-trifluoromethoxyphenyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydropropionic acid was obtained.

HPLC-MS (Method B): m / z = 744 (M + l); R _t = 9. 35 min.

(R) -3- {4- [3- [3- (tert-butyldimethylsilanyloxymethyl) -4-trifluoromethoxyphenyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino } -2-hydropropionic acid (17 mg, 0.023 mmol) was dissolved in acetonitrile: water (9: 1) (2 mL) and cesium fluoride (35 mg, 0.35 mmol) was added. The reaction mixture was stirred at 80 ° C. for 6 hours and an additional amount of cesium fluoride (35 mg) was added. The mixture was stirred overnight at 60 ° C., then concentrated in vacuo and diluted with ethyl acetate (10 mL) and water (5 mL). The organic phase was washed with water (3 x 5 mL), dried over magnesium sulfate and concentrated in vacuo. The residue was purified by preparative HPLC to give the title compound .

¹ H-NMR (DMSO- d ₆ ): δ 1.35 (m, 5H), 1.79 (m, 5H), 4.49 (s, 2H), 4.95 (s, 2H), 5.29 (s, 1H), 7. 12-7.26 (m, 6H), 7.34 (d, 2H), 7.49 (dd, 1H), 7.63 (d, 1H), 7.77 (d, 2H), 8.42 (t, 1H); HPLC MS (Method B): m / z = 630 (M + l); R _t = 6.62 min.

General method (C) for solid phase synthesis of a compound of formula (Ic) :

In the above formula

R ² , R ³ , R ⁴ , R ⁵ , A, Z, D and E are as defined in formula (I),

X is -C (O) NH- or -C (O) NHCR ¹² R ^13- , wherein R ¹² and R ¹³ are as defined in formula (I),

Resin is a polystyrene resin loaded with 2-chlorotrityl linker.

Step 4 is performed using 1) BSA and 2) DN = C═O or D—CHR ¹³ —N═C═O when A is —CHOH—.

Otherwise step 4 is performed using only DN = C = O or D-CHR ¹³ -N = C = O.

The method is described in the following example.

Example 40 (General Method (C))

(R) -3- {4- [1- (4-tert-butylphenyl) -3- (3,4-dichlorophenyl) ureidomethyl] benzoylamino} -2-hydropropionic acid

Step 1: Resin Binding (R) -Fmoc-Isoserine

50 mg polystyrene resin functionalized with 2-chlorotrityl chloride was vortexed with N-methyl-2-pyrrolidinone (500 μL) and 1,2-dichloropropane (500 μL) for 1 hour. The resin was filtered off and washed with N-methyl-2-pyrrolidinone: 1,2-dichloropropane (1: 1,2 × 1 mL). N-methyl-2-pyrrolidinone (500, μL) and 1,2-dichloropropane (500 μL) were added followed by 150 μmol (R) -Fmoc-isoserine and 100 μL diisopropylethylamine. . After shaking the suspension at 25 ° C. for 4 hours, the resin was separated by filtration, DCM: Methanol: Diisopropylethylamine 17: 2: 1 (2 × 1 mL) and N-methyl-2-pyrrolidinone (2 x 1 mL).

Step 2: Resin Bond (R) -3- (4-formylbenzoylamino) -2-hydroxypropionic acid

500 μL of a 20% solution of piperidine in DMF was added to the resin binding (R) -Fmoc-isoserine. After shaking for 30 minutes, the resin was drained and washed with N-methyl-2-pyrrolidinone (6 × 1 mL). Then 200 μmol 4-formylbenzoic acid (30 mg) and 200 μmol HOBt (31 mg) were dissolved in N-methyl-2-pyrrolidinone (500 μL) and added to the resin followed by acetonitrile (500 μL). Dissolved 200 μmol diisopropyl carbodimid (25.2 mg) was added. The mixture was shaken at 25 ° C. for 4 h, then filtered and the resin washed with N-methyl-2-pyrrolidinone (3 × 1 mL).

Step 3: Resin Bond (R) -3- {4-[(4-tert-butylphenylamino) methyl] benzoylamino} -2-hydroxypropionic acid

4- the resin bond (R) -3- (4-formylbenzoylamino) -2-hydropropionic acid in a mixture of N-methyl-2-pyrrolidinone and trimethylautoformate (1: 1,0.5 mL) 0.5 M solution of tert-butylaniline (0.25 mmol) and cold acetic acid (50 μL) were treated at 25 ° C. for 1 hour. Sodium cyanoborohydride (250 μmol, 16 mg) dissolved in N-methyl-2-pyrrolidinone and methanol (1: 1,0.25 mL) was added and the mixture was vortexed at 25 ° C. for 4 hours. Filtered to N-methyl-2-pyrrolidinone and methanol (1: 1,2 x 1 mL), 3 x 1 mL N-methyl-2-pyrrolidinone (3 x 1 mL), and 1,2-dichloro Wash with a mixture of propane and diisopropylethylamine (7: 1,2 x 0.75 mL).

Step 4: Resin Bond (R) -3- {4- [1- (4-tert-butylphenyl) -3- (3,4-dichlorophenyl) ureidomethyl] -benzoylamino} -2-hydropropionic acid

1,2-dichloropropane (500 μL) and BSA (100 μL) were added to the resin bond (R) -3- {4-[(4-tert-butylphenylamino) methyl] benzoylamino} -2-hydropropionic acid. Was added and the mixture was vortexed at 25 ° C. for 1 h. 200 μmol 3,4-dichlorophenylisocyanate is added and the mixture is shaken at 25 ° C. for 5 hours and then filtered and 2 × 1 mL DCM, 4 × 1 mL N-methyl-2-pyrrolidinone, 2 × 1 mL H _The resin was triturated with ₂ 0, 3 × 1 mL THF and 5 × 1 mL DCM to give the resin bound title compound .

Step 5: (R) -3- {4- [1- (4-tert-butylphenyl) -3- (3,4-dichlorophenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

The resin bond (R) -3- {4- [1- (4-tert-butylphenyl) -3- (3,4-dichlorophenyl) ureidomethyl] -benzoylamino} -2-hydropropionic acid for 1 hour Was treated with 1 mL 20% TFA in DCM at 25 ° C. The product was filtered off and the resin washed with 1 mL DCM. The combined extracts were concentrated in vacuo to afford the title compound .

¹ H-NMR (CDCl ₃ ): δ7.65 (d, 2H), 7.45-7.40 (m, 4H), 7.35-7.20 (m, 3H), 7.10-7.00 (m, 3H), 6.30 (s, 1H ), 4.90 (s, 2H), 4.40 (m, 1H), 3.83 (m, 2H), 1.32 (s, 9H); HPLC-MS (Method B): m / z = 558 (M + l); R _t = 4.71 min.

The following example is prepared as described above.

Example 41 (General Method (C))

(R) -3- {4- [1- (4-tert-butylcyclohexyl) -3- (3,4-dichlorophenyl) ureidomethyl] benzoylamino} -2 hydropropionic acid

HPLC-MS (Method B): m / z = 564 (M + l); R _t = 4.92 min / 5. 02 min.

Example 42 (General Method (C))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (3,4-dichlorophenyl) ureidomethyl] benzoylamino} -2-hydropropionic acid

HPLC-MS (Method B): m / z = 584 (M + l); R _t = 5.12 min.

Example 43 (General Method (C))

(R) -3- {4- [l- (4-cyclohexylphenyl) -3- (2,2,4,4-tetrafluoro-4 H -Benzo [1,3] dioxin-6-yl) ureidomethyl] benzoylamino} -2-hydropropionic acid

HPLC-MS (Method B): m / z = 646 (M + l); R _t = 5.24 min.

Example 44 (General Method (C))

(R) -3- {4- [1- (4-tert-butylphenyl) -3- (2,2,4,4-tetrafluoro-4 H -Benzo [1,3] dioxin-6-yl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

HPLC-MS (Method B): m / z = 620 (M + l); R _t = 4.88 min.

Example 45 (General Method (C))

(R) -3- {4- [1- (4-tert-butylcyclohexyl) -3- (2,2,4,4-tetrafluoro-4H-benzo [1,3] dioxine-6- Yl) ureidomethyl] benzoylamino} -2-hydropropionic acid

HPLC-MS (Method B): m / z = 606 (M + l); R _t = 5.11 min / 5. 20 min.

Example 46 (General Method (C))

(R) -3- {4- [1- (4-tert-butylphenyl) -3- (3,4-difluorophenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

HPLC-MS (Method B): m / z = 526 (M + l); Rt = 4.24 min.

Example 47 (General Method (C))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (3,4-difluorophenyl) ureidomethyl] benzoylamino} -2 hydropropionic acid

HPLC-MS (Method B): m / z = 552 (M + l); R _t = 4.65 min.

Example 48 (General Method (C))

(R) -3- {4- [1- (4-cyclohex-1-enylphenyl) -3- (3,4-difluorophenyl) ureidomethyl] benzoylamino} -2 hydropropionic acid

HPLC-MS (Method A): m / z = 550 (M + l); R _t = 6.77 min.

Example 49 (General Method (C))

(R) -3- {4- [3- (4-chloro-3-trifluoromethylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydroxypropionic acid

HPLC-MS (Method A): m / z = 618 (M + l); R _t = 7.58 min.

Example 50 (General Method (C))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (4-fluoro-3-nitrophenyl) ureidomethyl] benzoylamino} -2 hydropropionic acid

HPLC-MS (Method A): m / z = 579 (M + l); R _t = 6.85 min.

Example 51 (General Method (C))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (4-isopropylphenyl) ureidomethyl] benzoylamino} -2 hydropropionic acid

HPLC-MS (Method A): m / z = 558 (M + l); R _t = 7.73 min.

Example 52 (General Method (C))

(R) -3- {4- [1- (4-cyclohex-1-enylphenyl) -3- (3,4-dichlorophenyl) ureidomethyl] benzoylamino} -2 hydropropionic acid

HPLC-MS (Method B): m / z = 582 (M + l); R _t = 4.99 min.

Example 53 (General Method (C))

(R) -3- {4- [3- (4-acetylphenyl) -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2-hydropropionic acid

HPLC-MS (Method A): m / z = 558 (M + l); R _t = 6.42 min.

Example 54 (General Method (C))

3- {4- [3- [1 (RS)-(4-Bromophenyl) ethyl] -1- (4-cyclohexylphenyl) ureidomethyl] benzoylamino} -2 (R) -hydroxypropionic acid

HPLC-MS (Method A): m / z = 624 (M + l); R _t = 7.45 min.

Example 55 (General Method (C))

(R) -3- {4- [1- (4-cyclohexylphenyl) -3- (3,5-difluorophenyl) ureidomethyl] benzoylamino} -2 hydropropionic acid

HPLC-MS (Method B): m / z = 552 (M + l); R _t = 4.76 min.

General method (D) for solid phase synthesis of a compound of formula (Id):

In the above formula

R ² , R ³ , R ⁴ , R ⁵ , A, Z, D and E are as defined in formula (I),

X is -C (O)-(CR ¹² R ¹³ ) _r- (CH ₂ ) _s -where r, s, R ¹² and R ¹³ are as defined in formula (I),

Resin is a polystyrene resin loaded with 2-chlorotrityl linker.

Step 4 is carried out using 1) BSA and 2) DC (O) OH or D-CHR ¹³ -C (O) OH when A is -CHOH-. Otherwise, step 4 is performed using only DC (O) OH or D-CHR ¹³ -C (O) OH.

Example 56 (General Method (D))

(R) -3- [4-({(4-tert-butylcyclohexyl)-[2- (4-trifluoromethoxyphenyl) acetyl] amino} methyl) benzoylamino] -2-hydroxypropionic acid

Step 1: Resin Binding (R) -Fmoc-Isoserine

50 mg polystyrene resin functionalized with 2-chlorotrityl chloride was vortexed with N-methyl-2-pyrrolidinone (500 μL) and 1,2-dichloropropane (500 μL) for 1 hour. The resin was filtered off and washed with N-methyl-2-pyrrolidinone: 1,2-dichloropropane (1: 1,2 × 1 mL). N-methyl-2-pyrrolidinone (500, μL) and 1,2-dichloropropane (500 μL) were added followed by 150 μmol (R) -Fmoc-isoserine and 100 μL diisopropylethylamine. . After shaking the suspension at 25 ° C. for 4 hours, the resin was separated by filtration, DCM: Methanol: Diisopropylethylamine 17: 2: 1 (2 × 1 mL) and N-methyl-2-pyrrolidinone (2 x 1 mL).

Step 2: Resin Bond (R) -3- (4-formylbenzoylamino) -2-hydroxypropionic acid

500 μL of a 20% solution of piperidine in DMF was added to the resin binding (R) -Fmoc-isoserine. After shaking for 30 minutes, the resin was drained and washed with N-methyl-2-pyrrolidinone (6 × 1 mL). 200 μmol 4-formylbenzoic acid (30 mg) and 200 μmol HOBt (31 mg) were then dissolved in N-methyl-2-pyrrolidinone (500 μL) and added to the resin followed by acetonitrile (500 μL). Dissolved 200 μmol diisopropyl carbodimid (25.2 mg) was added. The mixture was shaken at 25 ° C. for 4 h, then filtered and the resin washed with N-methyl-2-pyrrolidinone (3 × 1 mL).

Step 3: Resin Bond (R) -3- {4-[(4-tert-Butylcyclohexylamino) methyl] benzoylamino} -2-hydroxypropionic acid

4- the resin bond (R) -3- (4-formylbenzoylamino) -2-hydropropionic acid in a mixture of N-methyl-2-pyrrolidinone and trimethylautoformate (1: 1,0.5 mL) Treatment with 0.5 M solution of tert-butylcyclohexylamine (0.25 mmol) and cold acetic acid (50 μL) at 25 ° C. for 1 hour. Sodium cyanoborohydride (250 μmol, 16 mg) dissolved in N-methyl-2-pyrrolidinone and methanol (1: 1,0.25 mL) was added and the mixture was vortexed at 25 ° C. for 4 hours. Filtered to N-methyl-2-pyrrolidinone and methanol (1: 1,2 x 1 mL), 3 x 1 mL N-methyl-2-pyrrolidinone (3 x 1 mL), and 1,2-dichloro Wash with a mixture of propane and diisopropylethylamine (7: 1,2 x 0.75 mL).

Step 4: Resin Bond (R) -3- [4-((4-tert-butylcyclohexyl)-[2- (4-trifluoromethoxyphenyl) acetyl] -amino} methyl) benzoylamino] -2- Hydropropionic acid

1,2-dichloropropane (500 μL) and BSA (100 μL) to the above resin bond (R) -3- {4-[(4-tert-butylcyclohexylamino) methyl] benzoylamino} -2-hydropropionic acid Was added and the mixture was vortexed at 25 ° C. for 1 h and then filtered. 4- (trifluoromethoxy) phenylacetic acid (400 μmol) in a mixture of N-methyl-2-pyrrolidinone, 1,2-dichloropropane and diisopropylethylamine (4.5: 4.5: 1,1 mL) to the resin ) Solution was added followed by bropo-tris (pyrrolidino) phosphonium hexafluorophosphate (400 μmol) in 1,2-dichloropropane (500 μL). The mixture was allowed to react at 50 ° C. for 3 hours and the resin was cooled to 25 ° C. while washing with N-methyl-2-pyrrolidinone (4 × 1 mL), and DCM (10 × 1 mL) to give the resin binding title The compound was obtained.

Step 5: (R) -3- [4-({(4-tert-butylcyclohexyl)-[4- (4-trifluoromethoxyphenyl) acetyl] amino} methyl) benzoylamino] -2-hydropropionic acid

Resin bond (R) -3- {4- [1- (4-tert-butylcyclohexyl) -3- (4-trifluoromethoxyphenyl) ureidomethyl] -benzoylamino} -2-hydropropionic acid Treated with 1 mL 20% TFA in DCM at 25 ° C. for 1 hour. The product was filtered off and the resin washed with 1 mL DCM. The combined extracts were concentrated in vacuo to afford the title compound .

HPLC-MS (Method A): m / z = 579 (M + l); R _t = 7.20 min.

The following example is prepared as described above.

Example 57 (General Method (D))

(R) -3- [4-({(4-tert-butylcyclohexyl)-[2- (3-fluoro-5-trifluoromethylphenyl) acetyl] amino} methyl) benzoylamino] -2-hydrate Roxypropionic acid

HPLC-MS (Method A): m / z = 581 (M + l); R _t = 7.22 min.

Example 58 (General Method (D))

(R) -3- [4-({(2,2-diphenylethyl)-[2- (3-fluoro-5-trifluoromethylphenyl) acetyl] amino} methyl) benzoylamino] -2-hydrate Roxypropionic acid

HPLC-MS (Method A): m / z = 623 (M + l); R _t = 6. 87 min.

Example 59 (General Method (D))

(R) -3- [4-{[(5-chlorobenzo [b] thiophen-3-carbonyl)-(2,2-diphenylethyl) amino] methyl} benzoylamino) -2-hydroxypropionic acid

HPLC-MS (Method A): m / z = 613 (M + l); R _t = 6.50 min.

Example 60 (General Method (D))

(R) -3- [4-({(2,2-diphenylethyl)-[2- (4-trifluoromethoxyphenyl) acetyl] amino} methyl) benzoylamino] -2-hydropropionic acid

HPLC-MS (Method A): m / z = 621 (M + l); R _t = 6.90 min.

Example 61 (General Method (D))

(R) -3- (4-{[(4-tert-butylcyclohexyl)-(5-chlorobenzo [b] thiophen-3-carbonyl) amino] methyl} benzoylamino) -2-hydroxypropionic acid

HPLC-MS (Method A): m / z = 571 (M + l); R _t = 7.15 min.

Example 62 (General Method (D))

(R) -3- (4-{[(2,2-diphenylethyl)-(5-trifluoromethoxy-1H-indole-2-carbonyl) amino] methyl} benzoylamino) -2-hydroxy Propionic acid

HPLC-MS (Method A): m / z = 646 (M + l); R _t = 6.93 min.

Example 63 (General Method (D))

(R) -3- [4-({(4-cyclohexylphenyl)-[(4-trifluoromethoxyphenyl) acetyl] amino} methyl) benzoylamino] -2-hydroxypropionic acid

HPLC-MS (Method B): m / z = 599 (M + l); R _t = 5.19 min.

Example 64 (General Method (D))

(R) -3- [4-({(4-cyclohexylphenyl)-[(3-trifluolmethoxyphenyl) acetyl] amino} methyl) benzoylamino] -2-hydroxypropionic acid

HPLC-MS (Method B): m / z = 599 (M + l); R _t = 5.17 min.

Example 65 (General Method (D))

(R) -3- [4-({(4-cyclohexylphenyl)-[(3-fluoro-5-trifluoromethylphenyl) acetyl] amino} methyl) benzoylaminol-2-hydropropionic acid

HPLC-MS (Method B): m / z = 601 (M + l); R _t = 5.19 min.

Example 66 (General Method (D))

(R) -3- (4-{([(3,5-Bis (trifluoromethyl) phenyl) acetyl]-(4-cyclohexylphenyl) amino) methyl} benzoylamino) -2-hydropropionic acid

HPLC-MS (Method B): m / z = 651 (M + l); R _t = 5.50 min.

Example 67 (General Method (D))

(R) -3- [4-({(4-cyclohexylphenyl)-[(3-trifluolmethylphenyl) acetyl] amino} methyl) benzoylamino] -2-hydroxypropionic acid

HPLC-MS (Method B): m / z = 583 (M + l); R _t = 5.08 min.

Example 68 (General Method (D))

(R) -3- [4-({(4-cyclohexylphenyl)-[(3,4-thichlorophenyl) acetyl] amino} methyl) benzoylamino] -2-hydropropionic acid

HPLC-MS (Method B): m / z = 583 (M + l); R _t = 5.26 min.

Example 69 (General Method (D))

(R) -3- (4-{[[(3-bromophenyl) acetyl]-(4-cyclohexylphenyl) amino] methyl} benzoylamino) -2-hydroxypropionic acid

HPLC-MS (Method B): m / z = 595 (M + l); R _t = 5.01 min.

Example 70 (General Method (D))

(R) -3- (4-{[(biphenyl-4-ylacetyl)-(4-cyclohexylphenyl) amino] methyl} benzoylamino) -2-hydroxypropionic acid

HPLC-MS (Method B): m / z = 591 (M + l); R _t = 5.38 min.

Example 71 (General Method (D))

(R) -3- (4-{[(4-cyclohexylphenyl)-(2-naphthylacetyl) amino] methyl} benzoylamino) -2-hydropropionic acid

HPLC-MS (Method B): m / z = 565 (M + l); R _t = 5.10 min.

Example 72 (General Method (D))

(R) -3- (4-{[(3- (3,5-Bis (trifluoromethyl) phenyl) propionyl)-(4-cyclohexylphenyl) amino] methyl} benzoylamino) -2-hydrate Roxypropionic acid

HPLC-MS (Method B): m / z = 665 (M + l); R _t = 5.51 min.

Example 73 (General Method (D))

(R) -3- [4-({(4-cyclohexylphenyl)-[3- (3-nitrophenyl) propionyl] amino} methyl) benzoylamino] -2-hydropropionic acid

HPLC-MS (Method B): m / z = 665 (M + l); R _t = 5.51 min.

The following preferred compounds are within the scope of the present invention and may be preferred according to the methods disclosed herein. Other preferred compounds are as follows:

In the above formula

Moreover, the following compounds are within the scope of the present invention and may be preferred according to the methods disclosed herein:

Furthermore, the following preferred compounds (as pure enantiomers of either (R) or (S) stereoconfiguration or mixtures thereof comprising racemates) are within the scope of the invention and described in the foregoing specification. May be preferred.

Pharmaceutical method

The following section describes binding assays as well as functional assays useful for evaluating the efficacy of the compounds of the present invention.

Binding of compounds to glucagon receptors can be measured in a competitive binding assay using cloned human glucagon receptors.

Antagonism can be measured as the ability of a compound to inhibit the amount of cAMP formed in the presence of 5 nM glucagon.

Glucagon binding assay (I)

Receptor binding was assayed using cloned human receptors (Lok et al., Gene 140,203-209 (1994)). Receptor (Lok et al.) Inserted into the pLJ6 'expression vector was expressed in a young hamster kidney cell line (A3 BHK 570-25) using the EcoR1 / SSt1 restriction enzyme site. Clones were selected in the presence of 0.5 mg / mL G-418, which appeared to be stable over 40 passages. K _d was found to be 0.1 nM.

Growing cells were isolated from the surface by cold buffer (30 mM NaCl, 1 mM dithiothreitol, 5 mg / L leupetin (Sigma), 5 mg / L pepstatin (Sigma), 100 mg / L bacitracin). (Sigma) and resuspended in 10 mM tris / HCI, pH 7.4 containing 15 mg / L recombinant aprotinin (Novo Nordisk A / S), and both 10 using Poltron PT 10-35 homogenizer (Kinematica) Prepare the plasma membrane by homogenizing by s rupture and centrifugation at 95.000 × g for 75 minutes in 41 w / v% sucrose. The white band located between the two layers is diluted with buffer and centrifuged at 40.000 x g for 45 minutes. Precipitates containing plasma membranes are suspended in buffer and stored at -80 ° C until use.

Glucagon is iodinated according to the chloramine T method (Hunter and Greenwood, Nature 194,495 (1962)) and purified using anion exchange chromatography (Jogensen et al., Hormone and Metab. Res. 4,223-224 (1972). Activity is 460 μCi / μg on the day of iodide Tracer is dispensed and stored at -18 ° C and used immediately after thawing.

Binding assays are performed on filter microtiter plates (MADV N65, Millipore). The buffer is 50 mM HEPES, 5 mM EGTA, 5 mM MgCl ₂ , 0.005% tween 20, pH 7.4. Glucagon is dissolved in 0.05 M HCI and lyophilized after addition of the same amount (w / w) of human serum albumin. On the day of use, dissolve in water and dilute to the desired concentration with buffer.

Test compounds are dissolved and diluted in DMSO. 140 μL buffer, 25 μL Klucagon or buffer, and 10 μL DMSO or test compound are added to each well. Dilute tracer (50.000 cpm) with buffer and add 25 μL to each well. 1-4 μg freshly thawed plasma membrane protein diluted in buffer is then added to each well in 25 μL aliquots. Plates are incubated at 30 ° C. for 2 hours. Non-specific binding is measured with ^10-6 M glucagon. The combined and unbound tracers are then separated by a vacuum manifold. Plates are washed with 2 x 100 μL buffer / well. The plate is allowed to air dry for 2 hours and the filter is separated from the plate using a Millipore Puncher. Count the filters in a gamma counter.

Function test (I)

Functional assays were performed in 96 well microtiter plates (tissue culture plates, Nunc). The resulting buffer concentrations in this assay are 50 mM tris / HCI, 1 mM EGTA, 1.5 mM MgSO ₄ , 1.7 mM ATP, 20 uM GTP, 2 mM IBMX, 0.02% tween-20 and 0.1% human serum albumin. pH is 7.4. Glucagon and the proposed antagonist were diluted in 50 mM tris / HCI, 1 mM EGTA, 1.85 mM MgS04, 0.0222% tween-20 and 0.111% human serum albumin, pH 7.4, and added in aliquots at 35 μL. 20 μL of 50 mM tris / HCI, 1 mM EGTA, 1.5 mM MgSO ₄ , 11.8 mM ATP, 0.14 mM GTP, 14 mM IBMX and 0.1% human serum albumin, pH 7.4 were added. GTP was dissolved just before the assay.

50 μL containing 5 μg plasma membrane protein is added to tris / HCI, EGTA, MgSO ₄ , human serum albumin buffer (actual concentration depends on the concentration of plasma membrane stored).

Total assay volume is 140 μL. The plate is incubated with shaking at 37 ° C. for 2 hours. The reaction was terminated by addition of 25 μL 0.5 N HCI. cAMP was measured using a scintillation proximity kit (Amersham).

Glucagon Binding Assay (II)

BHK (young hamster kidney cell line) cells are transfected with human glucagon receptor and membrane specimens of the cells are prepared. SPA beads (WGA beads) derived from Wheat Germ Agglutinin containing scintillant (Amersham) are bound to the membrane. ¹²⁵ I-glucagon binds to the human glucagon receptor on the membrane and excites the scintillation in the WGA to luminesce. Glucagon or sample bound to the receptor competes with ¹²⁵ I-glucagon.

All steps of membrane sampling are carried out on ice or at 4 ° C. BHK cells are harvested and centrifuged. Pellets are resuspended in homogenation buffer (25 mM HEPES, pH = 7.4, 2.5 mM CaI ₂ , 1.0 mM MgCl ₂ , 250 mg / L bacitracin, 0.1 mM Pefabloc) and Poltron 10-35 homogena Homogenize with 2 x 10 seconds using Kinematica and add the same amount of homogenation buffer used for resuspension. After centrifugation (2000 xg for 15 minutes), the supernatant is transferred to a cold centrifuge tube and centrifuged at 40.000 xg for 45 minutes. The pellet is resuspended in homogenation buffer, homogenized to 2 x 10 seconds (Polytron) and additional homogenization buffer is added. The suspension is centrifuged at 40.000 xg for 45 minutes and the pellet is resuspended in resuspension buffer (25 mM HEPES, pH = 7.4,2.5 mM CaI ₂ , 1.0 mM MgCl ₂ ) and homogenized at 2 x 10 seconds. Polytron. Protein concentrations are usually around 1.75 mg / mL. Stabilization buffer (25 mM HEPES, pH = 7.4, 2.5 mM CaCl ₂ , 1.0 mM MgCl ₂ , 1% bovine serum albumin, 500 mg / L bacitracin, 2.5 M sutroose) was added and the membrane specimen was -80 Store at ℃.

Glucagon binding assays are performed on Optistyrene Microplates (Packard). 50 μL Assay Buffer (25 mM HEPES, pH = 7.5, 2.5 mM CaCl ₂ , 1.0 mM MgCl ₂ , 0.003% Tween-20, 0.005% Bacitracin, 0.05% Sodium Azide) and 5 μL Glucagon or Test Compound ( In DMSO) is added to each well. Then 50 μL membrane (7.5 ug) containing 50 μL tracer ( ¹²⁵ I-porcine glucagon, 50.000 cpm) and human glucagon receptor is added to the wells. Finally, 50 μL WGA beads containing 1 mg beads are transferred to the wells. Opti plates are allowed to stand for 8 to 48 hours after incubation in shaker for 4 hours. Count the OptiPlate from the Topcounter. Non-specific binding is measured with 500 nM glucagon.

The compounds according to the examples showed IC50 values of 1500 nM or less and many of the compounds showed 250 nM or less when tested in the glucagon binding assay (II).

GIP binding assay

BHK (young hamster kidney cell line) cells are transfected with human GIP receptors and membrane specimens of the cells are prepared. SPA beads (WGA beads) derived from Wheat Germ Agglutinin containing scintillant (Amersham) are bound to the membrane. ¹²⁵ I-GIP binds to the human GIP receptor on the membrane and excites the scintillator in the WGA beads to cause luminescence. The GIP or sample bound to the receptor competes with ¹²⁵ I-GIP.

All steps of membrane sampling are carried out on ice or at 4 ° C. BHK cells are harvested and centrifuged. Pellets are resuspended in homogenation buffer (25 mM HEPES, pH = 7.4, 2.5 mM CaI ₂ , 1.0 mM MgCl ₂ , 250 mg / L bacitracin, 0.1 mM Pefabloc) and Poltron 10-35 homogena Homogenize with 2 x 10 seconds using Kinematica and add the same amount of homogenation buffer used for resuspension. After centrifugation (2000 xg for 15 minutes), the supernatant is transferred to a cold centrifuge tube and centrifuged at 40.000 xg for 45 minutes. The pellet is resuspended in homogenation buffer, homogenized to 2 x 10 seconds (Polytron) and additional homogenization buffer is added. The suspension is centrifuged at 40.000 xg for 45 minutes and the pellet is resuspended in resuspension buffer (25 mM HEPES, pH = 7.4,2.5 mM CaI ₂ , 1.0 mM MgCl ₂ ) and homogenized at 2 x 10 seconds. Polytron. Protein concentrations are usually around 1.75 mg / mL. Stabilization buffer (25 mM HEPES, pH = 7.4, 2.5 mM CaCl ₂ , 1.0 mM MgCl ₂ , 1% bovine serum albumin, 500 mg / L bacitracin, 2.5 M sutroose) was added and the membrane specimen was -80 Store at ℃.

GIP binding assays are performed on Optistyrene Microplates (Packard). 50 μL Assay Buffer (25 mM HEPES, pH = 7.5, 2.5 mM CaCl ₂ , 1.0 mM MgCl ₂ , 0.003% Tween-20, 0.005% Bacitracin, 0.05% Sodium Azide) and 5 μL GIP or Test Compound ( In DMSO) is added to each well. Then 50 μL membrane (20 ug) containing 50 μL tracer ( ¹²⁵ I-Forcin Glucagon, 50.000 cpm) and human GIP receptor is added to the wells. Finally, 50 μL WGA beads containing 1 mg beads are transferred to the wells. Opti plates are allowed to stand for 8 to 48 hours after incubation in shaker for 3.5 hours. Count the OptiPlate from the Topcounter. Non-specific binding is measured by 500 nM GIP.

Claims (71)

Compounds of formula (I), as well as any optical or geometric isomers or tautomers or mixtures thereof or pharmaceutically acceptable salts thereof: Formula I In the above formula R ¹ , R ² , R ³ , R ⁴ and R ⁵ are independently hydrogen or C _1-6 -alkyl, A is -C (O)-, -CH (OR⁶)- Or -CHF-, Wherein R ⁶ is hydrogen or C _1-6 -alkyl, Z is a divalent radical derived from arylene or a 5 or 6 membered heteroaromatic ring containing 1 or 2 heteroatoms selected from nitrogen, oxygen and sulfur, This _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 9, -NR 9 R 10 and C _1-6 - optionally with one or two groups selected from alkyl R ⁷ and R ⁸ Can be substituted, Wherein R ⁹ and R ¹⁰ are independently hydrogen or C _1-6 -alkyl, X is But here r is 0 or 1, q and s are independently 0, 1, 2 or 3, R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are independently hydrogen, C _1-6 -alkyl or C _3-8 -cycloalkyl, D is From here R ¹⁵ , R ¹⁶ R ¹⁷ and R ¹⁸ are independently; Hydrogen, halogen, -CN, -CHF ₂ , -CF ₃ , -OCF ₃ , -OCHF ₂ , -OCH ₂ CF ₃ , -OCF ₂ CHF ₂ , -S (O) ₂ CF ₃ , -SCF ₃ ,- NO ₂ , -OR ²¹ , -NR ²¹ R ²² , -SR ²¹ , -NR ²¹ S (O) ₂ R ²² , -S (O) ₂ NR ²¹ R ²² , -S (O) NR ²¹ R ²² ,- S (O) R ²¹ , -S (O) ₂ R ²¹ , -C (O) NR ²¹ R ²² , -OC (O) NR ²¹ R ²² , -NR ²¹ C (O) R ²² , -CH ₂ C (O) NR ²¹ R ²² , -OCH ₂ C (O) NR ²¹ R ²² , -OC (O) R ²¹ , -C (O) R ²¹ or -C (O) OR ²¹ , C _1-6 -alkyl, C _2-6 -alkenyl or C _2-6 -alkynyl, (Which is optionally _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 21, -NR 21 R 22 and C _1-6 - may be substituted with one or more substituents selected from alkyl) C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl, heterocyclyl, C _3-8 -cycloalkyl-C _1-6 -alkyl, C _3-8 -cycloalkyl-C _1-6- Alkoxy, C _3-8 -cycloalkyloxy, C _3-8 -cycloalkyl-C _1-6 -alkylthio, C _3-8 -cycloalkylthio, C _3-8 -cycloalkyl-C _2-6 -al alkenyl, C _3-8 - cycloalkyl, -C _2-6 - alkynyl, C _4-8 - cycloalkenyl -C _1-6 - alkyl, C _4-8 - cycloalkenyl -C _2-6 - alkenyl , C _4-8 - cycloalkenyl -C _2-6 - alkynyl, heterocyclyl, -C _1-6 - alkyl, heterocyclyl, -C _2-6 - alkenyl, heterocyclyl, -C _2-6 - Alkynyl, aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-Ci _-6 -alkoxy, aryl-C _1-6 -alkyl, aryl-C _2-6 -alkenyl, aryl-C _2-6 -Alkynyl, heteroaryl, heteroaryl-C _1-6 -alkyl, heteroaryl-C _2-6 -alkenyl or heteroaryl-C _2-6 -alkynyl, Wherein the ring portion is optionally selected from halogen, —C (O) OR ²¹ , —CN, —CF ₃ , —OCF ₃ , —N0 ₂ , —OR ²¹ , —NR ²¹ R ²² and C _1-6 -alkyl May be substituted with one or more substituents, Wherein R ²¹ and R ²² are independently hydrogen, C _1-6 -alkyl, aryl-C _1-6 -alkyl or aryl, Or R ²¹ and R ²² optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally comprise one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; Can form a 3 to 8 membered heterocyclic ring, Or when two of the groups R ¹⁵ to R ¹⁸ are located together at adjacent points, they may form a crosslinking-(CR ²³ R ²⁴ ) _a -O- (CR ²⁵ R ²⁶ ) _c -O- From here a is 0, 1 or 2, c is 1 or 2, R ²³ , R ²⁴ , R ²⁵ and R ²⁶ are independently hydrogen, C _1-6 -alkyl or fluorine, R ¹⁹ and R ²⁰ are independently hydrogen, C _1-6 -alkyl, C _3-8 -cycloalkyl or C _3-8 -cycloalkyl-C _1-6 -alkyl, E is But here R ²⁷ and R ²⁸ are independently Hydrogen, halogen, -CN, -CF ₃ , -OR ³² , -NR ³² R ³³ , C _1-6 -alkyl, C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl or aryl, Wherein the aryl group may be optionally substituted with one or more substituents selected from halogen, —CN, —CF ₃ , —NO ₂ , —OR ³² , —NR ³² R ³³ and C _1-6 -alkyl, Wherein R ³² and R ³³ are independently hydrogen or C _1-6 -alkyl, R ³² and R ³³ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally comprise one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; To form a 3 to 8 membered heterocyclic ring, R ²⁹ , R ³⁰ and R ³¹ are independently; Hydrogen, halogen, -CHF ₂ , -CF ₃ , -OCF ₃ , -OCHF ₂ , -OCH ₂ CF ₃ , -OCF ₂ CHF ₂ , -SCF ₃ , -OR ³⁴ , -NR ³⁴ R ³⁵ , -SR ³⁴ , -S (O) R ³⁴ , -S (O) ₂ R ³⁴ , -C (O) NR ³⁴ R ³⁵ , -OC (O) NR ³⁴ R ³⁵ , -NR ³⁴ C (O) R ³⁵ , -OCH ₂ C (O) NR ³⁴ R ³⁵ , -C (O) R ³⁴ or -C (O) OR ³⁴ , C _1-6 -alkyl, C _2-6 -alkenyl or C _2-6 -alkynyl, (It may be optionally substituted with one or more substituents selected from halogen, —CN, —CF ₃ , —OCF ₃ , —N0 ₂ , —OR ³⁴ , —NR ³⁴ R ³⁵ and C _1-6 -alkyl) C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl, heterocyclyl, C _3-8 -cycloalkyl-C _1-6 -alkyl, C _3-8 -cycloalkyl-C _2-6- alkenyl, C _3-8 - cycloalkyl, -C _2-6 - alkynyl, C _4-8 - cycloalkenyl -C _1-6 - alkyl, C _4-8 - cycloalkenyl -C _2-6 - Al alkenyl, C _4-8 - cycloalkenyl -C _2-6 - alkynyl, heterocyclyl, -C _1-6 - alkyl, heterocyclyl, -C _2-6 - alkenyl, heterocyclyl, -C _2-6 -Alkynyl, aryl, aryloxy, aroyl, aryl-C _1-6 -alkoxy, aryl-C _1-6 -alkyl, aryl-C _2-6 -alkenyl, aryl-C _2-6 -alkynyl, Heteroaryl, heteroaryl-C _1-6 -alkyl, heteroaryl-C _2-6 -alkenyl or heteroaryl-C _2-6 -alkynyl, Wherein the ring moiety is halogen, -CN, -CF ₃ , -OCF ₃ , -NO ₂ , -OR ³⁴ , -NR ³⁴ R ³⁵ and Optionally substituted with one or more substituents selected from C _1-6 -alkyl, Wherein R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl, Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur when optionally attached to the same nitrogen atom together with said nitrogen atom and optionally comprise one or two double bonds; To form a 3 to 8 membered heterocyclic ring containing, Or the radicals —O— (CH ₂ ) _t —CR ³⁶ R ³⁷ — (CH ₂ ) ₁ —O when two of the groups R ²⁹ , R ³⁰ and R ³¹ are attached together to a carbon atom of the same ring or to a carbon atom of a different ring _{-, - (CH 2) t} -CR 36 R 37 - (CH 2) l - , or ^{_{-S- (CH 2) t -CR 36}} R 37 - (CH 2) l can form a -S-, From here t and l are independently 0,1,2,3,4 or 5, R ³⁶ and R ³⁷ are independently hydrogen or C _1-6 -alkyl. A compound according to claim ¹ , wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are hydrogen. The compound according to claim 1 or 2, wherein A is -CHF-. The compound of claim 1 or 2, wherein A is -CH (OR⁶R, here R⁶Is as defined in claim 1. 5. Compounds according to claim 4, wherein A is -CH (OH)-. In any of the preceding terms, Z is And wherein R ⁷ and R ⁸ are as defined in claim 1. 7. The compound of claim 6 wherein Z is The compound characterized by the above-mentioned. The compound of any one of the preceding claims, wherein X is Wherein q is 0 or 1, r is 0 or 1, s is 0, 1 or 2 and R ¹² and R ¹³ are independently hydrogen or C _1-6 -alkyl. The compound of claim 8, wherein X is —C (O) NH—, —C (O) NHCH ₂ —, —C (O) NHCH (CH ₃ ) —, —C (O) NHC (CH ₃ ) ₂ —, -C (O) NHCH ₂ CH _2- , -C (O) CH _2- , C (O) CH ₂ CH _2- , -C (O) CH = CH-,-(CH ₂ ) _s- , -C (O)-, -C (O) O- or -NHC (O)-, wherein s is 0 or 1. The compound of claim 9, wherein X is —C (O) NH—, —C (O) NHCH ₂ —, —C (O) NHCH (CH ₃ ) —, —C (O) NHCH ₂ CH ₂ —, —C (O) CH _2- , -C (O) CH = CH-,-(CH ₂ ) _s- , -C (O)-, -C (O) O- or -NHC (O)-, wherein s is 0 or 1; The compound of claim 10, wherein X is —C (O) NH—, —C (O) NHCH ₂ —, —C (O) NHCH (CH ₃ ) —, —C (O) NHCH ₂ CH ₂ —, —C (O) CH _2- , -CH _2- , -C (O)-or -NHC (O)-. The compound of claim 11, wherein X is —C (O) NH—, —C (O) NHCH ₂ —, —C (O) NHCH (CH ₃ ) —, —C (O) CH ₂ — or —C (O). )-. 13. Compounds according to claim 12, wherein X is -C (O) NH-. The compound of any one of the preceding claims, wherein D is And wherein R ¹⁵ , R ¹⁶ , R ¹⁷ , R ¹⁸ , R ¹⁹ and R ²⁰ are as defined in claim 1. 15. The method of claim 14 wherein D is And wherein R ¹⁵ , R ¹⁶ and R ¹⁷ are as defined in claim 1. The compound of claim 14 or 15, wherein R ¹⁵ , R ¹⁶ and R ¹⁷ are independently hydrogen, halogen, —CN, —NO ₂ , —CF ₃ , —OCF ₃ , —SCF ₃ , C _1-6 -alkyl , C _1-6 -alkoxy, -SC _1-6 -alkyl, -C (O) OR ²¹ , -C (O) R ²¹ , -CH ₂ OR ²¹ , -C (O) NR ²¹ R ²² , -S (O) R ²¹ , -S (O) ₂ R ²¹ , -S (O) ₂ CF ₃ , -S (O) ₂ NR ²¹ R ²² , C _3-8 -cycloalkyl, C _3-8 -cycloalkyl -C _{1-6 -alcohol} or C _3-8 -cycloalkyl-C _1-6 -thioalkyl, or Is aryl, heteroaryl or aryloxy which may be optionally substituted with -CF ₃ , -OCF ₃ , C _1-6 -alkyl, halogen or -C (O) OR ²¹ , or When two of R ¹⁵ , R ¹⁶ and R ¹⁷ are located together at adjacent points they form a crosslinking-(CR ²³ R ²⁴ ) _a -O- (CR ²⁵ R ²⁶ ) _c -O-, Wherein R ²¹ and R ²² are independently hydrogen or C _1-6 -alkyl and a, c, R ²³ , R ²⁴ , R ²⁵ and R ²⁶ are as defined in claim 1. 18. The compound of claim 16, wherein R ¹⁵ , R ¹⁶ and R ¹⁷ are independently hydrogen, halogen, -CN, -CF ₃ , -OCF ₃ or C _1-6 -alkoxy or R ¹⁵ , R ¹⁶ crosslinked together -CF To form ₂ -O-CF ₂ -O- and R ¹⁷ is hydrogen. 18. The compound of claim 17, wherein R ¹⁵ , R ¹⁶ and R ¹⁷ are independently hydrogen, halogen, -CN, -CF ₃ , -OCF ₃ or C _1-6 -alkoxy. 15. The method of claim 14 wherein D is And wherein R ¹⁵ , R ¹⁶ , R ¹⁹ and R ²⁰ are as defined in claim 1. 20. The compound of claim 19 wherein D is Wherein R ¹⁵ and R ¹⁶ are both hydrogen and R ¹⁹ is C _1-6 -alkyl, C _3-8 -cycloalkyl or C _3-8 -cycloalkyl-C _1-6 -alkyl compound. 20. The compound of claim 19 wherein D is And wherein R ¹⁵ and R ¹⁶ are both hydrogen and R ¹⁹ and R ²⁰ are both C _1-6 -alkyl. The compound of any one of the preceding claims, wherein E is And wherein R ²⁷ , R ²⁸ , R ²⁹ , R ³⁰ and R ³¹ are as defined in claim 1. 23. The compound of claim 22 wherein E is And wherein R ²⁷ and R ²⁸ are as defined in claim 1. The method of claim 23 wherein E is And wherein R ²⁷ and R ²⁸ are as defined in claim 1. The compound of claim 23 or 24, wherein R ²⁷ and R ²⁸ are independently hydrogen, C _1-6 -alkyl, C _3-8 -cycloalkyl, C _4-8 -cycloalkenyl or phenyl, wherein a phenyl group Is optionally substituted as defined in claim 1. The compound of claim 25, wherein R ²⁷ and R ²⁸ are independently hydrogen, C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl. ^27. The compound of claim 26, wherein R ²⁷ is hydrogen and R ²⁸ is C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl. ^28. The compound of claim 27, wherein R ²⁷ is hydrogen and R ²⁸ is C _1-6 -alkyl or C _3-8 -cycloalkyl. 23. The compound of claim 22 wherein E is And wherein R ²⁹ , R ³⁰ and R ³¹ are as defined in claim 1. The method of claim 29 wherein E is And wherein R ²⁹ , R ³⁰ and R ³¹ are as defined in claim 1. The compound of claim 29 or 30, wherein R ²⁹ , R ³⁰ and R ³¹ are independently And _{hydrogen, -CHF 2, -CF 3, -OCF} 3, -OCHF 2, -OCH 2 CF 3, -OCF 2 CHF 2, -SCF 3, -OR 34, -NR 34 R 35, -SR 34, - S (O) R ³⁴ , -S (O) ₂ R ³⁴ , -C (O) NR ³⁴ R ³⁵ , -OC (O) NR ³⁴ R ³⁵ , -NR ³⁴ C (O) R ³⁵ , -OCH ₂ C (O) NR ³⁴ R ³⁵ , -C (O) R ³⁴ or -C (O) OR ³⁴ , C _1-6 -ylkyl, C _2-6 -alkenyl or C _2-6 -alkynyl, (Which is a _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - may be optionally substituted with one or more substituents selected from alkyl) C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl, This _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - can be optionally substituted with one or more substituents selected from alkyl, Wherein R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl, Or when R ³⁴ and R ³⁵ are attached together with the nitrogen atom to the same nitrogen atom, optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally comprise one or two double bonds Compound which is capable of forming a 3 to 8 membered heterocyclic ring. The compound of claim 31, wherein R ²⁹ , R ^30, and R ³¹ are independently Hydrogen, C _1-6 -alkoxy, halogen, —CF ₃ , —OCF ₃ or R ³⁴ and R ³⁵ are —NR ³⁴ R ³⁵ as defined in claim 1, or C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl, optionally substituted as defined in claim 1. 33. The compound of claim 32, wherein R ²⁹ , R ³⁰ and R ³¹ are independently Hydrogen or C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl, optionally substituted as defined in claim 1. 33. The compound of claim 32, wherein R ²⁹ , R ³⁰ and R ³¹ are independently Hydrogen or C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl, And which _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - can be optionally substituted with one or more substituents selected from alkyl, Where R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl, Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; Compound which can form a 3 to 8 membered heterocyclic ring comprising a. 35. The compound of claim 34, wherein R ²⁹ and R ³¹ are both hydrogen and R ³⁰ is different from hydrogen. The compound of claim 34, wherein R ²⁹ and R ³¹ are both hydrogen and R ³⁰ is C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl, And which _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - can be optionally substituted with one or more substituents selected from alkyl, Where R ³⁴ and R ³⁵ are independently hydrogen, C 1-6 -alkyl or aryl, Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; Compound which can form a 3 to 8 membered heterocyclic ring comprising a. The compound of claim 36, wherein R ²⁹ and R ³¹ are both hydrogen and R ³⁰ is C _4-8 -cycloalkenyl, And which _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - can be optionally substituted with one or more substituents selected from alkyl, Where R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl, Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; Compound which can form a 3 to 8 membered heterocyclic ring comprising a. 38. The compound of claim 37, wherein R ²⁹ and R ³¹ are both hydrogen and R ³⁰ is cyclohexenyl, And which _{halogen, -CN, -CF 3, -OCF 3} , -NO 2, -OR 34, -NR 34 R 35 and C _1-6 - can be optionally substituted with one or more substituents selected from alkyl, Where R ³⁴ and R ³⁵ are independently hydrogen, C _1-6 -alkyl or aryl, Or R ³⁴ and R ³⁵ optionally further comprise one or two heteroatoms selected from nitrogen, oxygen and sulfur and optionally one or two double bonds when attached to the same nitrogen atom together with said nitrogen atom; Compound which can form a 3 to 8 membered heterocyclic ring comprising a. 39. The compound of claim 37 or 38, wherein R ³⁰ is substituted with one C _1-6 -alkyl substituent. 34. The compound of claim 33, wherein R ²⁹ , R ³⁰ and R ³¹ are independently hydrogen, C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl. 41. The compound of claim 40, wherein R ²⁹ and R ³¹ are both hydrogen and R ³⁰ is C _1-6 -alkyl, C _3-8 -cycloalkyl or C _4-8 -cycloalkenyl. A compound of formula I1 according to claim 1 Formula I1 Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , X, D and E are as defined in claim 1 or any of the preceding claims. A compound of formula I2 according to claim 1 Formula I2 Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , D and E are as defined in claim 1 or any of the preceding claims. A compound of formula I3 according to claim 1 Formula I3 Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ¹⁵ , R ¹⁶ , R ¹⁷ , R ²⁹ , R ³⁰ and R ³¹ are as defined in claim 1 or the preceding. As defined in any one of the clauses. 45. The compound of claim 42 or 43 or 44, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ and R ⁸ are hydrogen. A compound of formula I4 according to claim 1 Formula I4 Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , X, D and E are as defined in claim 1 or any of the preceding claims. A compound of formula I5 according to claim 1 Formula I5 Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁷ , R ⁸ , D and E are as defined in any one of claims 1 or preceding. 48. The compound of claim 46 or 47, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁷ and R ⁸ are hydrogen. The compound of any one of the preceding claims, having an IC ₅₀ value of no greater than 5 μM as measured as the glucagon binding assay (I) or glucagon binding assay (II) disclosed herein. 50. The method according to claim 49, having an IC ₅₀ value of less than 1 μM, preferably less than 500 nM and more preferably less than 100 nM as measured as the glucagon binding assay (I) or glucagon binding assay (II) disclosed herein. Compound. The compound according to any one of the preceding claims, which is a medicament useful for the treatment and / or prevention of indications selected from the group consisting of hyperglycemia, IGT, type 2 diabetes, type 1 diabetes, dyslipidemia and obesity. 52. The compound of any of claims 1-51, which is used as a drug. 52. A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 51 as an active ingredient together with one or more pharmaceutically acceptable carriers or excipients. The method of any one of claims 1-51, wherein from about 0.05 mg to about 1000 mg, preferably from about 0.1 mg to about 500 mg, particularly preferably from about 0.5 mg to about 200 mg in unit dosage form. A pharmaceutical composition comprising a compound that follows. Use of a compound according to any one of claims 1 to 51 for the manufacture of a medicament for the treatment and / or prevention of anomalies or diseases in which glucagon is beneficial. Use of a compound according to any one of claims 1 to 51 for the manufacture of a medicament for the treatment and / or prevention of glucagon-mediated abnormalities and diseases. Use of a compound according to any one of claims 1 to 51 for the manufacture of a medicament for the treatment and / or prevention of hyperglycemia. Use of a compound according to any one of claims 1 to 51 for the manufacture of a medicament for lowering blood sugar in a mammal. Use of a compound according to any one of claims 1 to 51 for the manufacture of a medicament for the treatment and / or prophylaxis of IGT. Use of a compound according to any one of claims 1 to 51 for the manufacture of a medicament for the treatment and / or prevention of type 2 diabetes. 61. The use according to claim 60, for the manufacture of a medicament for delaying or preventing the progression from IGT to type 2 diabetes. 61. The use according to claim 60, for the manufacture of a medicament for delaying or preventing the progression from non-insulin demanding type 2 diabetes to insulin demanding type 2 diabetes. Use of a compound according to any one of claims 1 to 51 for the manufacture of a medicament for the treatment and / or prevention of type 1 diabetes. Use of a compound according to any one of claims 1 to 51 for the manufacture of a medicament for the treatment and / or prevention of obesity. Use of a compound according to any one of claims 1 to 51 for the manufacture of a medicament for the treatment and / or prevention of dyslipidemia. 66. The use according to any one of claims 55 to 65, wherein the regimen further comprises treatment with an antidiabetic agent. 67. The use of any of claims 55 to 66, wherein the regimen further comprises treatment with an anti-obesity agent. 68. The use according to any one of claims 55 to 67 further comprising treatment with further antihyperlipidemic agents. 70. The use according to any one of claims 55 to 68 further comprising treatment with further antihypertensive agents. 52. An abnormal condition in which glucagon has an antagonistic action comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 51 or a pharmaceutical composition according to claim 53 or 54 or Methods of treating and / or preventing diseases. The method of claim 70, wherein the effective amount of the compound is in the range of about 0.05 mg to about 2000 mg, preferably about 0.1 mg to about 1000 mg and particularly preferably about 0.5 mg to about 500 mg per day.