Selective estrogen receptor degraders and uses thereof

Abstract

The present disclosure provides compounds of Formula (I) and Formula (II). The compounds described herein may be useful in treating proliferative diseases (e.g., cancer). Also provided in the present disclosure are pharmaceutical compositions, kits, methods, and uses including or using a compound described herein. ##STR00001##

Claims

1. A compound of Formula (I): ##STR00105## or a pharmaceutically acceptable salt thereof, wherein: A is —CR.sup.A═ or —N═, as valency permits; W is —NH—, —O—, or —S—; a is 1, 2, or 3; n is 1, 2, 3, or 4; each instance of R.sup.1 is independently hydrogen, halogen, substituted or unsubstituted alkyl, —OR.sup.A, or —CN; R.sup.2 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl; R.sup.3 is hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, —OR.sup.A, or —N(R.sup.B).sub.2; R.sup.4 is hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl, —OR.sup.A or —N(R.sup.B).sub.2; R.sup.5 is fluorine or chlorine; R.sup.6 is chlorine; R.sup.7 is hydrogen, halogen, substituted or unsubstituted alkyl, —OR.sup.A or —N(R.sup.B).sub.2; R.sup.A is hydrogen or substituted or unsubstituted alkyl, or oxygen protecting group; and R.sup.B is hydrogen or substituted or unsubstituted alkyl, nitrogen protecting group, or optionally two R.sup.B are taken together with the intervening atoms to form substituted or unsubstituted heterocyclyl or substituted or unsubstituted heteroaryl.

2. A compound of the formula: ##STR00106## ##STR00107## ##STR00108## or or a pharmaceutically acceptable salt thereof.

3. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

4. A method of treating breast cancer, the method comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition of claim 3.

5. The method of claim 4, wherein the breast cancer is ER+ breast cancer.

6. The compound of claim 1, wherein the compound is of the formula (IA): ##STR00109## or a pharmaceutically acceptable salt thereof.

7. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein A is —CH═.

8. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein W is —NH—.

9. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein a is 1.

10. The compound of claim 9, or a pharmaceutically acceptable salt thereof, wherein R.sup.1 is —OMe, —CF.sub.3, or —CN.

11. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein n is 1.

12. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.2 is methyl or —CF.sub.3.

13. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.3 is fluorine or methyl.

14. The compound of claim 13, or a pharmaceutically acceptable salt thereof, wherein R.sup.4 is fluorine or methyl.

15. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R.sup.7 is —CH.sub.2F.

16. A pharmaceutical composition comprising a compound of claim 2, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

17. A method of treating breast cancer, the method comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition of claim 16.

18. The method of claim 17, wherein the breast cancer is ER+ breast cancer.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) Non-limiting embodiments of the present disclosure will be described by way of example with reference to the accompanying figures, which are schematic and are not intended to be drawn to scale. In the figures, each identical or nearly identical component illustrated is typically represented by a single numeral. For purposes of clarity, not every component is labeled in every figure, nor is every component of each embodiment of the invention shown where illustration is not necessary to allow those of ordinary skill in the art to understand the invention.

(2) FIG. 1 is a chart showing the tumor volume change in a human breast cancer xMCF-7 xenograph efficacy study in a mouse model, under treatment by exemplary compound 25, compared with fulvestrant, GDC-0810 and AZD9496.

(3) FIG. 2 is a chart showing the tumor volume change in a human breast cancer xMCF-7 xenograph efficacy study in a mouse model, under treatment by exemplary compounds 12, 21 and 25, compared with AZD9496.

(4) FIG. 3 is a chart showing the effect of treatment with a combination of exemplary compound 25 with CDK4/6 inhibitor palbocilib in the tumor volume change in a human breast cancer xMCF-7 xenograft xenograph efficacy study in a mouse model.

DETAILED DESCRIPTION

(5) The present disclosure provides Selective Estrogen Receptor Degrader (SERD) compounds, for example, the compounds of Formula (I), which selectively binds an estrogen receptor and lead to the degradation of the receptor. The compounds described herein are useful in reducing the level of an estrogen receptor (wildtype or mutated) and treating a disease associated with a steroid hormone such as estrogen (e.g., breast cancer such as ER+ breast cancer). For example, the exemplary SERD compounds disclosed herein successfully induce degradation of ER and inhibit the growth of MCF-7 cancer cell, an ER+ human breast cancer cell line. Also provided in the present disclosure are pharmaceutical compositions, kits, methods of using the SERD compounds described herein for treating any of the target diseases described herein.

(6) Selective Estrogen Receptor Degraders

(7) One aspect of the present disclosure relates to the SERD compounds as described herein, as well as their pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, or prodrugs. These compounds are useful in treating and/or preventing proliferative diseases (such as ER+ breast cancer) or diseases associated with ER in a subject.

(8) In certain embodiments, a compound described herein is of Formula (I):

(9) ##STR00025##
in which R.sup.1-R.sup.7, A, W, a, and n are as described herein, or pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

(10) In Formula (I), A and W are in a tricyclic ring. In some embodiments, A can be —CR.sup.A═ or —N═. In some embodiments, A can be —CR.sup.A═, in which R.sup.A is as defined herein. In some embodiments, R.sup.A can be hydrogen. In some embodiments, R.sup.A can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In one example, A can be —CH═. In another example, A can be —CMe=. In another embodiments, A can be —NH═.

(11) In some embodiments, W can be —NH—. In some embodiments, W can be —O—. In some embodiments, W can be —S—.

(12) Formula (I) includes one or more instances of R.sup.1. In some embodiments, a can be 1. In some embodiments, a can be 2. In some embodiments, a can be 3. In some embodiments, at least one instance of R.sup.1 can be hydrogen. In some embodiments, at least one instance of R.sup.1 can be halogen (e.g., F, Cl, Br, or I). In some embodiments, at least one instance of R.sup.1 can be fluorine. In some embodiments, at least one instance of R.sup.1 can be chlorine. In some embodiments, at least one instance of R.sup.1 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In some embodiments, at least one instance of R.sup.1 can be —CF.sub.3. In some embodiments, at least one instance of R.sup.1 can be —OR.sup.A, in which R.sup.A is as defined herein. In some embodiments, at least one instance of R.sup.1 can be —OMe. In some embodiments, at least one instance of R.sup.1 can be —OEt. In some embodiments, at least one instance of R.sup.1 can be —CN.

(13) In Formula (I), in some embodiments, R.sup.2 can be hydrogen. In some embodiments, R.sup.2 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In some embodiments, R.sup.2 can be methyl. In some embodiments, R.sup.2 can be ethyl. In some embodiments, R.sup.2 can be propyl. In some embodiments, R.sup.2 can be —CF.sub.3. In some embodiments, R.sup.2 can be substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system). In some embodiments, R.sup.2 can be cyclopropyl. In some embodiments, R.sup.2 can be cyclobutyl.

(14) In Formula (I), in some embodiments, R.sup.3 and/or R.sup.4 can be hydrogen. In some embodiments, R.sup.3 and/or R.sup.4 can be halogen (e.g., F, Cl, Br, or I). In some embodiments, R.sup.3 and/or R.sup.4 can be fluorine. In some embodiments, R.sup.3 and/or R.sup.4 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In some embodiments, R.sup.3 and/or R.sup.4 can be methyl. In some embodiments, R.sup.3 and/or R.sup.4 can be ethyl. In some embodiments, R.sup.3 and/or R.sup.4 can be propyl. In some embodiments, one of R.sup.3 and R.sup.4 can be fluorine, and the other of R.sup.3 and R.sup.4 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl). In some embodiments, one of R.sup.3 and R.sup.4 can be fluorine, and the other of R.sup.3 and R.sup.4 can be methyl. In some embodiments, both R.sup.3 and R.sup.4 can be fluorine. In some embodiments, both R.sup.3 and R.sup.4 can be methyl. In some embodiments, R.sup.3 and/or R.sup.4 can be substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system). In some embodiments, R.sup.3 and/or R.sup.4 can be substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 5- to 10-membered monocyclic or bicyclic heterocyclic ring, wherein one or two atoms in the heterocyclic ring are independently nitrogen, oxygen, or sulfur). In some embodiments, R.sup.3 and/or R.sup.4 can be substituted or unsubstituted aryl (e.g., phenyl, or benzyl). In some embodiments, R.sup.3 and/or R.sup.4 can be substituted or unsubstituted 5- to 7-membered monocyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur. In some embodiments, R.sup.3 and/or R.sup.4 can be —OR.sup.A, in which R.sup.A is as defined herein (e.g., —OH, —O(substituted or unsubstituted C.sub.1-6 alkyl) (e.g., —OMe)). In some embodiments, R.sup.3 and/or R.sup.4 can be —N(R.sup.B).sub.2, in which R.sup.B is as defined herein (e.g., —NH.sub.2).

(15) In Formula (I), in some embodiments, R.sup.5 and/or R.sup.6 can be hydrogen. In some embodiments, R.sup.5 and/or R.sup.6 can be halogen (e.g., F, Cl, Br, or I). In some embodiments, R.sup.5 and/or R.sup.6 can be chlorine. In some embodiments, R.sup.5 and/or R.sup.6 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In some embodiments, R.sup.5 and R.sup.6 can both be halogen. In some embodiments, R.sup.5 and R.sup.6 can both be fluorine. In some embodiments, R.sup.5 and R.sup.6 can both be chlorine. In some embodiments, one of R.sup.5 and R.sup.6 can be chlorine, and the other one can be fluorine. In some embodiments, one of R.sup.5 and R.sup.6 can be fluorine, and the other one can be substituted or unsubstituted C.sub.1-6 alkyl. In some embodiments, one of R.sup.5 and R.sup.6 can be chlorine, and the other one can be substituted or unsubstituted C.sub.1-6 alkyl.

(16) In Formula (I), in some embodiments, R.sup.7 can be hydrogen. In some embodiments, R.sup.7 can be halogen (e.g., F, Cl, Br, or I). In some embodiments, R.sup.7 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In some embodiments, R.sup.7 can be of the formula: —CH.sub.2R.sup.a, in which R.sup.a is halogen. In some embodiments, R.sup.7 can be —CH.sub.2F. In some embodiments, R.sup.7 can be —OR.sup.A, in which R.sup.A is as defined herein (e.g., —OH, —O (substituted or unsubstituted C.sub.1-6 alkyl) (e.g., —OMe)). In some embodiments, R.sup.7 can be —N(R.sup.B).sub.2, in which R.sup.B is as defined herein (e.g., —NH.sub.2).

(17) In Formula (I), in some embodiments, n can be 1. In some embodiments, n can be 2. In some embodiments, n can be 3. In some embodiments, n can be 4.

(18) In certain embodiments, a compound described herein is of Formula (II):

(19) ##STR00026##
in which R.sup.1-R.sup.4, R.sup.8, R.sup.A1, R.sup.A2, A, W, and a are as described herein, or pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

(20) In Formula (II), A and W are in a tricyclic ring. In some embodiments, A can be —CR.sup.A═ or —N═. In some embodiments, A can be —CR.sup.A═, in which R.sup.A is as defined herein. In some embodiments, R.sup.A can be hydrogen. In some embodiments, R.sup.A can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In one example, A can be —CH═. In one example, A can be —CMe=. In another embodiments, A can be —NH═.

(21) In some embodiments, W can be —NH—. In some embodiments, W can be —O—. In some embodiments, W can be —S—.

(22) Formula (II) includes one or more instances of R.sup.1. In some embodiments, a can be 1. In some embodiments, a can be 2. In some embodiments, a can be 3. In some embodiments, at least one instance of R.sup.1 can be hydrogen. In some embodiments, at least one instance of R.sup.1 can be halogen (e.g., F, Cl, Br, or I). In some embodiments, at least one instance of R.sup.1 can be fluorine. In some embodiments, at least one instance of R.sup.1 can be chlorine. In some embodiments, at least one instance of R.sup.1 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In some embodiments, at least one instance of R.sup.1 can be substituted or unsubstituted methyl. In some embodiments, at least one instance of R.sup.1 can be —CF.sub.3. In some embodiments, at least one instance of R.sup.1 can be substituted or unsubstituted ethyl. In some embodiments, at least one instance of R.sup.1 can be substituted or unsubstituted propyl. In some embodiments, at least one instance of R.sup.1 can be —OR.sup.A, in which R.sup.A is as defined herein. In some embodiments, at least one instance of R.sup.1 can be —OH. In some embodiments, at least one instance of R.sup.A can be substituted or unsubstituted C.sub.1-6 alkyl. In some embodiments, at least one instance of R.sup.1 can be —OMe. In some embodiments, at least one instance of R.sup.1 can be —CN.

(23) In Formula (II), in some embodiments, R.sup.2 can be hydrogen. In some embodiments, R.sup.2 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In some embodiments, R.sup.2 can be substituted or unsubstituted methyl. In some embodiments, R.sup.2 can be —CF.sub.3. In some embodiments, R.sup.2 can be substituted or unsubstituted ethyl. In some embodiments, R.sup.2 can be substituted or unsubstituted propyl. In some embodiments, R.sup.2 can be substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system). In some embodiments, R.sup.2 can be substituted or unsubstituted cyclopropyl.

(24) In Formula (II), in some embodiments, R.sup.3 and/or R.sup.4 can be hydrogen. In some embodiments, R.sup.3 and/or R.sup.4 can be halogen (e.g., F, Cl, Br, or I). In some embodiments, R.sup.3 and/or R.sup.4 can be fluorine. In some embodiments, R.sup.3 and R.sup.4 can both be fluorine. In some embodiments, R.sup.3 and/or R.sup.4 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In some embodiments, R.sup.3 and/or R.sup.4 can be methyl. In some embodiments, one of R.sup.3 and R.sup.4 can be fluorine, and the other can be methyl. In some embodiments, one of R.sup.3 and R.sup.4 can be fluorine, and the other can be hydrogen. In some embodiments, one of R.sup.3 and R.sup.4 can be methyl, and the other can be hydrogen. In some embodiments, R.sup.3 and/or R.sup.4 can be substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system). In some embodiments, R.sup.3 and/or R.sup.4 can be substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 5- to 10-membered monocyclic or bicyclic heterocyclic ring, wherein one or two atoms in the heterocyclic ring are independently nitrogen, oxygen, or sulfur). In some embodiments, R.sup.3 and/or R.sup.4 can be substituted or unsubstituted aryl (e.g., phenyl, or benzyl). In some embodiments, R.sup.3 and/or R.sup.4 can be substituted or unsubstituted 5- to 7-membered monocyclic heteroaryl, wherein one, two, three, or four atoms in the heteroaryl ring system are independently nitrogen, oxygen, or sulfur. In some embodiments, R.sup.3 and/or R.sup.4 can be —OR.sup.A, in which R.sup.A is as defined herein (e.g., —OH, —O(substituted or unsubstituted C.sub.1-6 alkyl) (e.g., —OMe)). In some embodiments, R.sup.3 and/or R.sup.4 can be —N(R.sup.B).sub.2, in which R.sup.B is as defined herein (e.g., —NH.sub.2). In some embodiments, R.sup.3 and R.sup.4 can be taken together with the intervening atoms to form substituted or unsubstituted carbocyclyl (e.g., substituted or unsubstituted, 3- to 7-membered, monocyclic carbocyclyl comprising zero, one, or two double bonds in the carbocyclic ring system). In some embodiments, R.sup.3 and R.sup.4 can be taken together with the intervening atoms to form substituted or unsubstituted cyclopropyl. In some embodiments, R.sup.3 and R.sup.4 can be taken together with the intervening atoms to form unsubstituted cyclopropyl, and R.sup.8 can be fluorine. In some embodiments, R.sup.3 and R.sup.4 can be taken together with the intervening atoms to form unsubstituted cyclopropyl, and R.sup.8 can be methyl. In some embodiments, R.sup.3 and R.sup.4 can be taken together with the intervening atoms to form substituted or unsubstituted heterocyclyl (e.g., substituted or unsubstituted, 5- to 10-membered monocyclic or bicyclic heterocyclic ring, wherein one or two atoms in the heterocyclic ring are independently nitrogen, oxygen, or sulfur).

(25) In Formula (II), in some embodiments, R.sup.8 can be hydrogen. In some embodiments, R.sup.8 can be halogen (e.g., F, Cl, Br, or I). In some embodiments, R.sup.8 can be fluorine. In some embodiments, R.sup.8 can be substituted or unsubstituted methyl. In some embodiments, R.sup.8 can be methyl.

(26) In Formula (II), in some embodiments, R.sup.A1 and/or R.sup.A2 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In some embodiments, R.sup.A1 and/or R.sup.A2 can be substituted or unsubstituted methyl. In some embodiments, R.sup.A1 and/or R.sup.A2 can be substituted or unsubstituted ethyl. In some embodiments, R.sup.A1 and/or R.sup.A2 can be substituted or unsubstituted propyl. In some embodiments, R.sup.A1 and/or R.sup.A2 can be chlorine. In some embodiments, R.sup.A1 and/or R.sup.A2 can be fluorine. In some embodiments, either R.sup.A1 or R.sup.A2 can be chlorine. In some embodiments, both R.sup.A1 and R.sup.A2 can be chlorine. In some embodiments, one of R.sup.A1 or R.sup.A2 can be fluorine, and the other one of R.sup.A1 and R.sup.A2 can be selected from the group consisting of substituted or unsubstituted alkyl, chlorine, and fluorine. In some embodiments, one of R.sup.A1 or R.sup.A2 can be fluorine, and the other one of R.sup.A1 and R.sup.A2 can be substituted or unsubstituted C.sub.1-6 alkyl (e.g., methyl, ethyl, or propyl). In some embodiments, one of R.sup.A1 or R.sup.A2 can be fluorine, and the other one of R.sup.A1 and R.sup.A2 can be methyl. In some embodiments, one of R.sup.A1 or R.sup.A2 can be fluorine, and the other one of R.sup.A1 and R.sup.A2 can be chlorine. In some embodiments, both R.sup.A1 and R.sup.A2 can be fluorine.

(27) In some embodiments, the compound of Formula (I) can be of the formula of the compounds described herein, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

(28) In some embodiments, the compound of Formula (II) can be of the formula of the compounds described herein, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.

(29) The compounds described herein can be prepared from readily available starting materials using methods known in the art. It will be appreciated that where typical or preferred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, and pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvents used, but such conditions can be determined by those skilled in the art by routine optimization procedures. The chemicals used in the above-described synthetic routes may include, for example, solvents, reagents, catalysts, and protecting group and deprotecting group reagents. The methods described above may also additionally include steps, either before or after the steps described specifically herein, to add or remove suitable protecting groups in order to ultimately allow synthesis of the compounds. In addition, various synthetic steps may be performed in an alternate sequence or order to give the desired compounds. Synthetic chemistry transformations and protecting group methodologies (protection and deprotection) useful in synthesizing applicable compounds are known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3.sup.rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) and subsequent editions thereof.

(30) The compounds of Formula (I) and Formula (II) provided herein can be prepared from readily available starting materials using the following general methods and procedures. Exemplary schematic illustrations for synthesizing the compounds of the invention described herein are provided below. Where typical or preferred process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvents used, but such conditions can be determined by those skilled in the art by routine optimization procedures.

(31) The compounds described herein, e.g., compounds of formula (I), may be prepared according to the general Scheme A. Compounds A1, are commercially available or can be synthesized via standard transformations known to those of ordinary skill in the art of organic/medicinal chemistry. Compounds A3 can be prepared by Pictet-Spengler reaction of A1 with aldehyde A2 (X═I, Br, or Cl). Alkylation of A3 with A4 could produce compound A5. Alternatively, A5 could be prepared by alkylation of A1 with A4, followed by Pictet-Spengler reaction of A6 with aldehyde A2. Copper or palladium promoted C—O bond formation of compound A5 with ethylene glycol could provide compound A7. Derivatization of the OH group of A7 to a leaving group, followed by displacement with amine A8, could afford (I).

(32) ##STR00027##

(33) The compounds of formula (II) may be prepared according to the general Scheme B. Compounds B3, are commercially available or can be assembled via standard transformations known to those of ordinary proficiency in the art of organic/medicinal chemistry. Compounds B2 could be prepared by alkylation of A1 with B1. Pictet-Spengler reaction of B2 with aldehyde B3 could produce B4. The saponification of B4 to (II) could be generally accomplished by the use of an alkali metal hydroxide in aqueous or mixed aqueous/organic solvents.

(34) ##STR00028##

(35) Alternatively, the compounds of formula (II) may be prepared according to the general Scheme C. Pictet-Spengler reaction of B2 with aldehyde C1 (X═I, Br, or Cl) could produce C2. Compounds C4 could be prepared by palladium mediated Heck reaction of C2 with C3. The saponification of C4 to (II) could be generally accomplished by the use of an alkali metal hydroxide in aqueous or mixed aqueous/organic solvents

(36) ##STR00029##
Pharmaceutical Compositions and Kits

(37) The present disclosure provides pharmaceutical compositions comprising a compound described herein, or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable excipient. In certain embodiments, a pharmaceutical composition described herein comprises a compound described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. The pharmaceutical compositions described herein are useful in treating and/or preventing proliferative diseases (e.g., ER+ breast cancer) or diseases associated with ER.

(38) In certain embodiments, the cell contacted with an effective amount of a compound or pharmaceutical composition described herein is in vitro. In certain embodiments, the contacted cell is ex vivo. In certain embodiments, the cell described herein is in vivo. In certain embodiments, the cell described herein is a malignant cell (e.g., malignant breast cancer cell).

(39) In certain embodiments, the compound described herein is provided in an effective amount in the pharmaceutical composition. In certain embodiments, the effective amount is a therapeutically effective amount (e.g., amount effective for treating a proliferative disease in a subject in need thereof). In certain embodiments, the proliferative disease is cancer, e.g., ER+ breast cancer. In certain embodiments, the effective amount is a prophylactically effective amount (e.g., amount effective for preventing a proliferative disease in a subject in need thereof and/or for keeping a subject in need thereof in remission of a proliferative disease).

(40) Pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. In general, such preparatory methods include bringing the compound described herein (i.e., the “active ingredient”) into association with a carrier or excipient, and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping, and/or packaging the product into a desired single- or multi-dose unit.

(41) Pharmaceutical compositions can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. A “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage, such as one-half or one-third of such a dosage.

(42) Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition described herein will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered. The composition may comprise between 0.1% and 100% (w/w) active ingredient.

(43) Pharmaceutically acceptable excipients used in the manufacture of provided pharmaceutical compositions include inert diluents, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and perfuming agents may also be present in the composition.

(44) Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredients, the liquid dosage forms may comprise inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. In certain embodiments for parenteral administration, the conjugates described herein are mixed with solubilizing agents such as Cremophor®, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof.

(45) Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, U.S.P., and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

(46) The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

(47) In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form may be accomplished by dissolving or suspending the drug in an oil vehicle.

(48) Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, (c) humectants such as glycerol, (d) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarding agents such as paraffin, (f) absorption accelerators such as quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, (h) absorbents such as kaolin and bentonite clay, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may include a buffering agent.

(49) Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art of pharmacology. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulating compositions which can be used include polymeric substances and waxes. Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

(50) The active ingredient can be in a micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings, and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active ingredient can be admixed with at least one inert diluent such as sucrose, lactose, or starch. Such dosage forms may comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may comprise buffering agents. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulating agents which can be used include polymeric substances and waxes.

(51) Although the descriptions of pharmaceutical compositions provided herein are mainly directed to pharmaceutical compositions which are suitable for administration to humans, such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with ordinary experimentation.

(52) The compounds provided herein are typically formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compositions described herein will be decided by a physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject or organism will depend upon a variety of factors including the disease being treated and the severity of the disorder; the activity of the specific active ingredient employed; the specific composition employed; the age, body weight, general health, sex, and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific active ingredient employed; the duration of the treatment; drugs used in combination or coincidental with the specific active ingredient employed; and like factors well known in the medical arts.

(53) Also encompassed by the disclosure are kits (e.g., pharmaceutical packs). The kits provided may comprise a pharmaceutical composition or compound described herein and a container (e.g., a vial, ampule, bottle, syringe, and/or dispenser package, or other suitable container). In some embodiments, provided kits may optionally further include a second container comprising a pharmaceutical excipient for dilution or suspension of a pharmaceutical composition or compound described herein. In some embodiments, the pharmaceutical composition or compound described herein provided in the first container and the second container are combined to form one unit dosage form.

(54) In certain embodiments, a kit described herein includes a first container comprising a compound or pharmaceutical composition described herein. In certain embodiments, a kit described herein is useful in treating a proliferative disease (e.g., ER+ breast cancer) in a subject in need thereof, and/or preventing a proliferative disease in a subject in need thereof. In some embodiments, the SERDs described herein are useful in treating diseases and/or disorders associated with a steroid hormone such as estrogen.

(55) In certain embodiments, a kit described herein further includes instructions for using the compound or pharmaceutical composition included in the kit. A kit described herein may also include information as required by a regulatory agency such as the U.S. Food and Drug Administration (FDA). In certain embodiments, the information included in the kits is prescribing information. In certain embodiments, the kits and instructions provide for treating a proliferative disease in a subject in need thereof, and/or preventing a proliferative disease in a subject in need thereof. A kit described herein may include one or more additional pharmaceutical agents described herein as a separate composition.

(56) Methods of Treatment

(57) As shown in the Examples below, exemplary SERD compounds described herein successfully induced degradation of ER and inhibited the growth of ER+ breast cancer cells and exhibited better human hepatocyte clearance than drug of the same class such as fulvestrant, as well as those currently in clinical trials such as GDC-0810 and AZD9496. In mouse studies, these compounds also showed superior pharmacokinetic profiles (e.g., clearance, half life, and AUC) than drugs currently in clinical uses or clinical trials.

(58) Accordingly, the present disclosure provides methods of treating a proliferative disease and/or a disease associated with a steroid hormone such as estrogen, in a subject in need thereof, the methods comprising administering to the subject an effective amount (e.g., therapeutically effective amount) of a compound, or pharmaceutical composition thereof, described herein.

(59) Another aspect of the present disclosure relates to methods of preventing proliferative disease in a subject in need thereof, the methods comprising administering to the subject an effective amount (e.g., prophylactically effective amount) of a compound, or pharmaceutical composition thereof, described herein.

(60) The compounds and pharmaceutical compositions described herein are useful in treating and/or preventing proliferative diseases. In certain embodiments, the proliferative disease is cancer. In certain embodiments, the proliferative disease is breast cancer. In certain embodiments, the proliferative disease is gynecological disease or cancer associated with ER such as cancer of the ovary, cervix or endometrium and breast cancer, particularly ER+ breast cancer.

(61) In certain embodiments, the method described herein further includes administering to the subject an additional pharmaceutical agent. In certain embodiments, the method described herein further includes contacting the biological sample with an additional pharmaceutical agent. In certain embodiments, the method described herein further includes contacting the tissue with an additional pharmaceutical agent. In certain embodiments, the method described herein further includes contacting the cell with an additional pharmaceutical agent. In certain embodiments, the method described herein further includes radiotherapy, immunotherapy, and/or transplantation (e.g., bone marrow transplantation).

(62) The compounds and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops). Specifically contemplated routes are oral administration, intravenous administration (e.g., systemic intravenous injection), regional administration via blood and/or lymph supply, and/or direct administration to an affected site. In general, the most appropriate route of administration will depend upon a variety of factors including the nature of the agent (e.g., its stability in the environment of the gastrointestinal tract), and/or the condition of the subject (e.g., whether the subject is able to tolerate oral administration).

(63) The exact amount of a compound required to achieve an effective amount will vary from subject to subject, depending, for example, on species, age, and general condition of a subject, severity of the side effects or disorder, identity of the particular compound, mode of administration, and the like. An effective amount may be included in a single dose (e.g., single oral dose) or multiple doses (e.g., multiple oral doses). In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample, tissue, or cell, any two doses of the multiple doses include different or substantially the same amounts of a compound described herein. In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample, tissue, or cell, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is three doses a day, two doses a day, one dose a day, one dose every other day, one dose every third day, or one dose every week. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is one dose per day. In certain embodiments, the frequency of administering the multiple doses to the subject or applying the multiple doses to the tissue or cell is two doses per day. In certain embodiments, when multiple doses are administered to a subject or applied to a biological sample, tissue, or cell, the duration between the first dose and last dose of the multiple doses is one day, two days, four days, one week, two weeks, three weeks, one month, two months, three months, four months, six months, nine months, one year, two years, three years, four years, five years, seven years, ten years, fifteen years, twenty years, or the lifetime of the subject, biological sample, tissue, or cell. In certain embodiments, the duration between the first dose and last dose of the multiple doses is three months, six months, or one year. In certain embodiments, the duration between the first dose and last dose of the multiple doses is the lifetime of the subject, biological sample, tissue, or cell. In certain embodiments, a dose (e.g., a single dose, or any dose of multiple doses) described herein includes independently between 1 mg and 3 mg, between 3 mg and 10 mg, between 10 mg and 30 mg, between 30 mg and 100 mg, between 100 mg and 300 mg, between 300 mg and 1,000 mg, or between 1 g and 10 g, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 3 mg and 10 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 10 mg and 30 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 30 mg and 100 mg, inclusive, of a compound described herein. In certain embodiments, a dose described herein includes independently between 100 mg and 300 mg, inclusive, of a compound as described herein. In certain embodiments, a dose described herein includes independently between 300 mg and 1000 mg, inclusive, of a compound described herein.

(64) Dose ranges as described herein provide guidance for the administration of provided pharmaceutical compositions to an adult. The amount to be administered to, for example, a child or an adolescent can be determined by a medical practitioner or person skilled in the art and can be lower or the same as that administered to an adult.

(65) A compound or composition, as described herein, can be administered in combination with one or more additional pharmaceutical agents (e.g., therapeutically and/or prophylactically active agents) useful in treating and/or preventing a proliferative disease. The compounds or compositions can be administered in combination with additional pharmaceutical agents that improve their activity (e.g., activity (e.g., potency and/or efficacy) in treating a proliferative disease in a subject in need thereof, and/or in preventing a proliferative disease in a subject in need thereof), improve bioavailability, improve safety, reduce drug resistance, reduce and/or modify metabolism, inhibit excretion, and/or modify distribution in a subject, biological sample, tissue, or cell. It will also be appreciated that the therapy employed may achieve a desired effect for the same disorder, and/or it may achieve different effects. In certain embodiments, a pharmaceutical composition described herein including a compound described herein and an additional pharmaceutical agent shows a synergistic effect that is absent in a pharmaceutical composition including one of the compound and the additional pharmaceutical agent, but not both.

(66) The compound or composition can be administered concurrently with, prior to, or subsequent to one or more additional pharmaceutical agents, which may be useful as, e.g., combination therapies in treating and/or preventing a proliferative disease. Pharmaceutical agents include therapeutically active agents. Pharmaceutical agents also include prophylactically active agents. Pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved for human or veterinary use by the U.S. Food and Drug Administration as provided in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, antibodies, small molecules linked to proteins such as antibodies, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells. In certain embodiments, the additional pharmaceutical agent is a pharmaceutical agent useful in treating a proliferative disease. In certain embodiments, the additional pharmaceutical agent is a pharmaceutical agent useful in preventing a proliferative disease. In certain embodiments, the additional pharmaceutical agent is a pharmaceutical agent approved by a regulatory agency (e.g., the US FDA) for treating and/or preventing a proliferative disease. Each additional pharmaceutical agent may be administered at a dose and/or on a time schedule determined for that pharmaceutical agent. The additional pharmaceutical agents may also be administered together with each other and/or with the compound or composition described herein in a single dose or administered separately in different doses. The particular combination to employ in a regimen will take into account compatibility of the compound described herein with the additional pharmaceutical agent(s) and/or the desired therapeutic and/or prophylactic effect to be achieved. In general, it is expected that the additional pharmaceutical agent(s) in combination be utilized at levels that do not exceed the levels at which they are utilized individually. In some embodiments, the levels utilized in combination will be lower than those utilized individually.

(67) In certain embodiments, the additional pharmaceutical agent is an anti-proliferative agent (e.g., anti-cancer agent, for example, an immune-oncology agents (e.g., anti-PD-1 antibody) or cells (e.g., CAR-T cells)). In certain embodiments, the additional pharmaceutical agent is an anti-angiogenesis agent, anti-inflammatory agent, immunosuppressant, anti-bacterial agent, anti-viral agent, cardiovascular agent, cholesterol-lowering agent, anti-diabetic agent, anti-allergic agent, pain-relieving agent, or a combination thereof. In certain embodiments, the compounds described herein or pharmaceutical compositions can be administered in combination with an anti-cancer therapy including, but not limited to, targeted therapy (e.g., mTOR signaling pathway inhibitor), cell therapy, surgery, radiation therapy, immunotherapy, and chemotherapy (e.g., docetaxel, doxorubicin).

(68) Without further elaboration, it is believed that one skilled in the art can, based on the above description, utilize the present invention to its fullest extent. The following specific embodiments are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever. All publications cited herein are incorporated by reference for the purposes or subject matter referenced herein.

EXAMPLES

(69) In order that the present disclosure may be more fully understood, the following examples are set forth. The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions, and methods provided herein and are not to be construed in any way as limiting their scope.

Example 1: Synthesis of Compound 1 and Derivatives Thereof

(70) ##STR00030## ##STR00031##

Step 1. Synthesis of 1-2

(71) A solution of (2R)-1-(1H-indol-3-yl)propan-2-amine (2 g, 11.48 mmol), acetic acid (1 mL) and 2,6-difluoro-4-iodobenzaldehyde (3 g, 11.19 mmol) in Toluene (20 mL) was stirred at 80° C. for 12 h. The resulting mixture was then cooled to room temperature and concentrated under vacuum. The residue was purified by a silica gel column with ethyl acetate/petroleum ether (1/100-1/10) as the eluent to afford the desired product (2 g, 42% yield).

Step 2. Synthesis of 1-3

(72) To a solution of 1-2 (2 g, 4.71 mmol) in 1,4-dioxane (20 mL) were added N,N-diisopropylethylamine (920 mg, 7.08 mmol) and 2,2-difluoropropyl trifluoromethanesulfonate (1.61 g, 7.06 mmol). The resulting solution was stirred at 100° C. for 12 h. The reaction was cooled down to room temperature and then quenched with water (50 mL). The mixture was extracted with ethyl acetate (50 mL×3). The organic phase was washed with brine (50 mL) and concentrated under vacuum to afford the desired product (1.5 g, crude).

Step 3. Synthesis of 1-4

(73) To a solution of 1-3 (500 mg, 1.00 mmol) in ethane-1,2-diol (5 mL) were added copper iodide (94.6 mg, 0.50 mmol), 1,10-phenanthroline (18 mg, 0.10 mmol) and cesium carbonate (649 mg, 1.99 mmol). The resulting mixture was then stirred at 100° C. for 2 h. After cooling to room temperature, the mixture was diluted with water (20 mL), and extracted with ethyl acetate (30 mL×3). The organic phase was washed with brine (20 mL) and concentrated under vacuum. The residue was purified by silica gel column eluting with ethyl acetate/petroleum ether (1/100-1/10) to afford the desired product (200 mg, 46% yield).

Step 4. Synthesis of 1-5

(74) To a solution of 1-4 (50 mg, 0.11 mmol) in tetrahydrofuran (2 mL), were added triethylamine (13.9 mg, 0.14 mmol) and 4-methylbenzene-1-sulfonyl chloride (26 mg, 0.14 mmol). The resulting mixture was then stirred at room temperature for 3 h. The mixture was diluted with water (10 mL), and extracted with ethyl acetate (10 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated under vacuum to afford the desired product (50 mg, 74% yield).

Step 5. Synthesis of 1

(75) A solution of 1-5 (50 mg, 0.08 mmol), 3-(fluoromethyl)azetidine (20.47 mg, 0.23 mmol), and cesium carbonate (41.5 mg, 0.13 mmol) in acetonitrile (2 mL) was stirred at 80° C. for 12 h. After cooling to room temperature, the mixture was diluted with water (10 mL), and extracted with ethyl acetate (10 mL×3). The organic phase was concentrated under vacuum. The residue was purified by silica gel column eluting with ethyl acetate/petroleum ether (1/100-1/5) to afford the desired product (19.7 mg, 46%). LCMS (ES, m/z): 508.4. .sup.1HNMR (300 MHz, CDCl.sub.3, ppm): δ 7.41 (d, J=6.9 Hz, 1H), 7.19-7.17 (d, J=7.2 Hz, 1H), 7.03-6.95 (m, 2H) 6.64-6.59 (m, 2H), 5.24 (s, 1H), 4.61 (d, J=3.9 Hz, 1H), 4.46 (d, J=3.9 Hz, 1H), 4.18-4.10 (m, 4H), 3.96-3.90 (m, 2H), 3.57-3.55 (m, 1H), 3.44-3.41 (m, 2H), 3.31-3.10 (m, 1H), 3.10-2.95 (m, 2H), 2.65-2.60 (m, 2H), 1.42 (t, J=18.9 Hz, 3H), 1.13 (d, J=6.6 Hz, 3H).

(76) Using the similar procedures described above, the following additional compounds of the invention were prepared.

(77) TABLE-US-00001 TABLE 1 Exemplary Derivatives of Compound 1 Exam- ple No. Compound [M + H].sup.+ HNMR 2 490.3 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.44 (d, J = 6.9 Hz, 1H), 7.22 (d, J = 7.2 Hz, 1H), 7.07- 6.97 (m, 2H), 6.64-6.59 (m, 2H), 5.27 (s, 1H), 4.16 (t, J = 5.4 Hz, 2H), 3.65-3.59 (m, 1H), 3.11-3.01 (m, 2H), 2.96 (t, J = 5.4 Hz, 2H), 2.75-2.61 (m, 6H), 2.00-1.85 (m, 4H), 1.45 (t, J = 18.6 Hz, 3H), 1.18 (d, J = 6.6 Hz, 3H). 3 540.3 .sup.1HNMR (300 MHz, CDCl.sub.3, ppm): δ 7.43 (d, J = 7.2 Hz, 1H), 7.11 (d, J = 8.1 Hz, 1H), 7.06- 6.95 (m, 2H), 6.62-6.57 9m, 2H), 5.25 (s, 1H), 4.15 (t, J = 5.4 Hz, 2H), 3.63-3.57 (m, 1H), 3.10-3.00 (m, 2H), 2.90 (t, J = 5.4 Hz, 2H), 2.69-2.56 (m, 6H), 1.69-1.68 (m, 4H), 1.52- 1.46 (m, 2H), 1.43 (t, J = 18.9 Hz, 3H), 1.16 (d, J = 6.6 Hz, 3H). 4 518.3 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.43 (d, J = 8.4 Hz, 1H), 7.19 (d, J = 7.2 Hz, 1H), 7.06- 6.96 (m, 2 H), 6.62-6.57 (m, 2H), 5.25 (s, 1H), 4.13 (t, J = 5.7 Hz, 2H), 3.66-3.56 (m, 1H), 3.13-2.96 (m, 4H), 2.85-2.8 1m, 4H), 2.69- 2.59 (m, 2H), 1.72-1.67 (m, 8H), 1.43 (t, J = 18.6 Hz, 3 H) 1.16 (d, J = 6.6 Hz, 3 H). 5 504.4 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.40 (d, J = 1.8 Hz, 1 H), 7.38-7.37 (m, 1 H), 7.19-6.94 (m, 2 H), 6.51 (d, J = 10.5 Hz, 2 H), 5.18 (s, 1 H), 4.55 (d, J = 5.4 Hz, 1 H), 4.39 (d, J = 5.4 Hz, 1 H), 4.00 (t, J = 6.3 Hz, 2 H), 3.70-3.58 (m, 1 H), 3.51 (t, J = 7.5 Hz, 2 H), 3.20 (t, J = 6.9. Hz, 2 H), 3.01-2.95 (m, 1 H), 2.90-2.84 (m, 4 H), 2.62-2.55 (m, 1 H), 2.45-2.37 (m, 1 H), 1.20-1.08 (m, 9 H). 6 486.4 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.40 (d, J = 1.8 Hz, 1 H), 7.38-7.37 (m, 1 H), 7.18- 6.94 (m, 2 H), 6.57 (d, J = 10.5 Hz, 2 H), 5.18 (s, 1 H), 4.12 (t, J = 5.4 Hz, 2 H), 3.70-3.58 (m, 1 H), 3.07-2.84 (m, 4 H), 2.70-2.55 (m, 5 H), 2.46-2.32 (m, 1 H), 1.84 (m, 4 H), 1.20- 1.08 (m, 9 H). 7 500.5 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ7.43- 7.40 (m, 1 H), 7.21-7.18 (m, 1H), 7.06-6.95 (m, 2H), 6.60-6.54 (m, 2H), 5.21 (s, 1H), 4.12 (t, J = 5.4 Hz, 2 H), 3.70-3.58 (m, 1 H), 3.07-2.80 (m, 4 H), 2.76-2.57 (m, 5 H), 2.46- 2.32 (m, 1 H), 1.65-1.62 (m, 4 H), 1.50-1.48 (m, 2 H), 1.20-1.08 (m, 9 H). 8 514.5 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.42 (d, J = 6.9 Hz, 1 H), 7.20 (d, J = 7.8 Hz, 1 H), 7.04-6.95 (m, 2 H), 6.56 (d, J = 10.8 Hz, 2 H), 5.20 (s, 1H), 4.12 (t, J = 5.4 Hz, 2 H), 3.73- 3.66 (m, 1 H), 3.09-2.90 (m, l H), 2.84-2.81 (m, 3H), 2.87-2.71 (m, 4 H), 2.61 (d, J = 14.7 Hz, 1H), 2.49-2.35 (m, 1 H), 1.69-1.64 (m, 8H), 1.23-1.11 (m, 9 H). 9 536.2 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm) δ 7.39 (d, J = 6.9 Hz, 1H), 7.17 (d, J = 6.3 Hz, 1H), 7.05 (s, 1H), 7.05-6.93 (m, 2H), 6.85 (s, 1H), 5.61 (s, 1H), 4.54 (d, J = 5.7 Hz, 1H), 4.38 (d, J = 5.4 Hz, 1 H), 3.98 (t, J = 5.1 Hz, 2 H), 3.85- 3.75 (m, 1H), 3.50 (t, J = 7.8 Hz, 2H), 3.21- 3.13 (m, 3H), 2.97 (t, J = 14.4 Hz, 1H), 2.86- 2.83 (m, 3H), 2.63 (d, J = 14.7 Hz, 1H), 2.37- 2.22 (m, 1H), 1.21-1.06 (m, 9 H). 10 0 540.4 .sup.1HNMR (300 MHz, CDCl.sub.3, ppm) δ 7.54-7.51 (m, 1 H), 7.26-7.20 (m, 2 H), 7.16-7.09 (m, 2 H), 6.97 (s, l H), 6.84 (s, l H), 5.66 (s, 1H), 4.62 (d, J = 7.2 Hz, 1 H), 4.46 (d, J = 7.2 Hz, 1 H), 4.02 (t, J = 4.5 Hz, 2 H), 3.78-3.69 (m, 1 H), 3.62 (s, 2 H), 3.28-3.05 (m, 4 H), 2.93 (s, 3H), 2.73-2.64 (m, 1H), 2.60-2.48 (m, 1H), 1.45 (t, J = 18.9 Hz, 3H), 1.16 (d, J = 6.3 Hz, 3H). 11 520.2 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.38 (dd, J.sub.1 = 6.3 Hz, J.sub.2 = 2.1 Hz, 1H), 7.16 (dd, J.sub.1 = 6.3 Hz, J.sub.2 = 2.1 Hz, 1 H), 6.98-6.93 (m, 2H) 6.83 (d, J = 2.1 Hz, 1 H), 6.64 (dd, J.sub.1 = 12.0 Hz, J.sub.2 = 2.1 Hz, 1 H), 5.35 (s, 1 H), 4.55 (d, J = 5.4 Hz, 1H), 4.39 (d, J = 5.4 Hz, 1 H), 3.99 (t, J = 5.4 Hz, 2 H), 3.80-3.72 (m, 1 H), 3.54-3.52 (m, 2 H), 3.20-3.10 (m, 3 H), 2.95-2.84 (m, 4 H), 2.60 (d, J = 14.4 Hz, 1 H), 2.40-2.25 (m, 1H), 1.14-1.06 (m, 9 H). 12 524.2 .sup.1HNMR (300 MHz, CDCl.sub.3, ppm): δ7.55-7.52 (m, 1 H), 7.45 (s, 1 H), 7.25-7.22 (m, 1 H), 7.16-7.09 (m, 2 H), 6.79 (t, J = 1.5 Hz, 1 H), 6.53 (dd, J = 3 Hz, J = 12 Hz, 1H), 5.41 (s, 1 H), 4.61 (d, J = 5.7 Hz, 1 H), 4.45(d, J = 5.7 Hz, 1 H), 3.95 (t, J = 5.4 Hz, 2 H), 3.74-3.67 (m, 1 H), 3.52(t, J = 7.5 Hz, 2 H), 3.18 (t, J = 6.9 Hz, 3 H), 3.12-3.04 (m, 1H), 2.85 (t, J = 5.4 Hz, 3 H), 2.70-2.53 (m, 2 H), 1.42 (t, J = 18.9 Hz, 3 H), 1.15(d, J = 6.6 Hz, 3 H).

Example 2: Synthesis of Compounds 13B and Derivatives Thereof

(78) ##STR00043## ##STR00044##

Step 1. Synthesis of 13-1

(79) To a solution of 5-fluoro-1H-indole-3-carbaldehyde (2.0 g, 12.3 mmol, 1.00 equiv) in toluene (20 mL) were added ammonium acetate (1.14 g, 14.8 mmol, 1.20 equiv) and nitroethane (24 mL) at room temperature. The resulting solution was stirred at 130° C. for 5 h. The solid was filtered out and the filter cake was washed with ethyl acetate (10 mL×3). The filtrate was concentrated under vacuum, and the residue was diluted with water (40 mL) and extracted with ethyl acetate (35 mL×3). The combined organic layer was washed with brine (40 mL×3), dried over anhydrous sodium sulfate and concentrated under vacuum to afford 5-fluoro-3-[(1Z)-2-nitroprop-1-en-1-yl]-1H-indole (2.23 g, 83%) as a yellow solid.

Step 2. Synthesis of 13-2

(80) To a solution of 5-fluoro-3-[(1Z)-2-nitroprop-1-en-1-yl]-1H-indole (2.0 g, 9.1 mmol, 1.00 equiv) in tetrahydrofuran (50 mL) was added lithium aluminum hydride (1.37 g, 36.0 mmol, 4.00 equiv) at 0° C. and the reaction mixture was stirred at 65° C. for 1 hour. After cooling to room temperature, the reaction was quenched by the addition of water (1.4 mL), sodium hydroxide aqueous solution (4.2 mL, 10%). The solid was filtered out, and the filter cake was washed with tetrahydrofuran (15 mL×3). The filtrate was concentrated under vacuum to afford 1-(5-fluoro-1H-indol-3-yl)propan-2-amine (1.8 g, 102%) as a brown oil.

Step 3. Synthesis of 13-3

(81) To a solution of 1-(5-fluoro-1H-indol-3-yl)propan-2-amine (1.0 g, 5.2 mmol, 1.00 equiv) in 1,4-dioxane (24 mL) were added N,N-diisopropylethylamine (1.01 g, 7.8 mmol, 1.50 equiv) and 2-fluoro-2-methylpropyl trifluoromethanesulfonate (1.28 g, 5.7 mmol, 1.10 equiv). The resulting solution was stirred at 70° C. overnight. The reaction was quenched by the addition of water (30 mL) and the mixture was extracted with ethyl acetate (30 mL×3). The combined organic layer was washed with brine (30 mL×3), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:4) as eluent to afford [1-(5-fluoro-1H-indol-3-yl)propan-2-yl](2-fluoro-2-methylpropyl)amine (610 mg, 44%) as a brown solid.

Step 4. Synthesis of 13-4

(82) To a solution of [1-(5-fluoro-1H-indol-3-yl)propan-2-yl](2-fluoro-2-methylpropyl)amine (510 mg, 1.92 mmol, 1.00 equiv) in toluene (20 mL) were added acetic acid (460 mg, 7.67 mmol, 4.00 equiv) and 2,6-difluoro-4-iodobenzaldehyde (514 mg, 1.92 mmol, 1.00 equiv). The resulting solution was stirred at 80° C. overnight. The reaction was quenched by the addition of water (15 mL) and the mixture was extracted with ethyl acetate (20 mL×3), the combined organic layer was washed with brine (20 mL×3), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:50) as eluent to afford 1-(2,6-difluoro-4-iodophenyl)-6-fluoro-2-(2-fluoro-2-methylpropyl)-3-methyl-1H,2H,3H,4H,9H-pyrido[3,4-b]indole (800 mg, 81%) as a light yellow solid.

Step 5. Synthesis of 13-5

(83) To a solution of 1-(2,6-difluoro-4-iodophenyl)-6-fluoro-2-(2-fluoro-2-methylpropyl)-3-methyl-1H,2H,3H,4H,9H-pyrido[3,4-b]indole (680 mg, 1.32 mmol, 1.00 equiv) in ethylene glycol (20 mL) under N.sub.2 protection were added cesium carbonate (859 mg, 2.64 mmol, 2.00 equiv) and cuprous iodide (126 mg, 0.66 mmol, 0.50 equiv) was added. The resulting solution was stirred overnight at 100° C. and the reaction was cooled to room temperature. Then the reaction was quenched by the addition of water (20 mL) and the mixture was extracted with ethyl acetate (30 mL×3). The combined organic layer was washed with brine (40 mL×3), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:1) as eluent to afford 2-[3,5-difluoro-4-[6-fluoro-2-(2-fluoro-2-methylpropyl)-3-methyl-1H,2H,3H,4H,9H-pyrido[3,4-b]indol-1-yl]phenoxy]ethan-1-ol (600 mg, 101%) as light yellow oil.

Step 6. Synthesis of 13-6

(84) To a solution of 2-[3,5-difluoro-4-[6-fluoro-2-(2-fluoro-2-methylpropyl)-3-methyl-1H,2H,3H,4H,9H-pyrido[3,4-b]indol-1-yl]phenoxy]ethan-1-ol (550 mg, 1.22 mmol, 1.00 equiv) in dichloromethane (16 mL) were added triethylamine (247 mg, 2.44 mmol, 2.00 equiv) and N,N-dimethylaminopyridine (15 mg, 0.12 mmol, 0.10 equiv) under N.sub.2. 4-methylbenzene-1-sulfonyl chloride (280 mg, 1.45 mmol, 1.20 equiv) was added at 0° C. The resulting solution was stirred at room temperature for 2 hours. The reaction was quenched by the addition of water (15 mL) and the mixture was extracted with dichloromethane (20 mL×3). The combined organic layer was washed with brine (30 mL×3), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:5) as eluent to afford 2-[3,5-difluoro-4-[6-fluoro-2-(2-fluoro-2-methylpropyl)-3-methyl-1H,2H,3H,4H,9H-pyrido[3,4-b]indol-1-yl]phenoxy]ethyl 4-methylbenzene-1-sulfonate (400 mg, 54%) as light yellow oil.

Step 7. Synthesis of 13

(85) To a solution of 2-[3,5-difluoro-4-[6-fluoro-2-(2-fluoro-2-methylpropyl)-3-methyl-1H,2H,3H,4H,9H-pyrido[3,4-b]indol-1-yl]phenoxy]ethyl 4-methylbenzene-1-sulfonate (300 mg, 0.49 mmol, 1.00 equiv) in acetonitrile (18 mL) was added cesium carbonate (1.13 g, 3.48 mmol, 7.00 equiv) under N.sub.2. 3-(fluoromethyl)azetidine trifluoroacetyl (462 mg, 2.48 mmol, 5.00 equiv) was then added at 0° C. The resulting solution was stirred at 80° C. overnight. The reaction mixture was cooled to room temperature and the reaction was quenched by the addition of water (20 mL), the mixture was extracted with ethyl acetate (20 mL×3). The combined organic layer was washed with brine (30 mL×3), dried over anhydrous sodium sulfate and concentrated under vacuum. The crude product was purified by prep-HPLC [Column, Xbridge, RP18, 19*150 mm; mobile phase, A: NH.sub.4CO.sub.3 (aq) (5 mmol/L), B: acetonitrile (70%-90% in 8 min); rate, 25 mL/min; Detector, 254 nm] to afford compound 111 (100 mg, 38%) as a white solid. Compound 111 was resolved by chiral preparative HPLC [Chiral Column, IA-3, 250 mm×20 mm, 5 um; mobile phase, A: hexane (91%), B: ethanol (9%); rate, 20 mL/min; Detector, 254 nm] to afford 13A (retention time: 13.91 min) and 13B (retention time: 22.37 min).

13A

(86) 13.9 mg, white solid. Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% DEA)/IPA; Gradient: 10% IPA; Detector: 254 nm], Retention time=7.22 min. LCMS (ES, m/z) [M+H]+: 522.40. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm) δ 7.13 (dd, J=4.5 Hz, J=8.7 Hz, 1H), 7.07 (dd, J=2.4 Hz, 6.9 Hz, 1H), 6.76 (td, J=2.4 Hz, 9.0 Hz, 1H), 6.58-6.53 (m, 2H), 5.19 (s, 1H), 4.58 (d, J=7.2 Hz, 1H), 4.42 (d, J=7.2 Hz, 1H), 4.01 (t, J=6.8 Hz, 2H), 3.71-3.65 (m, 1H), 3.55 (t, J=10.0 Hz, 2H), 3.24 (t, J=10.0 Hz, 2H), 3.06-2.99 (m, 1H), 2.91-2.86 (m, 3H), 2.57 (dd, J=2.7 Hz, J=14.7 Hz, 1H), 2.40 (dd, J=15.0 Hz, J=26.4 Hz, 1H), 1.23-1.10 (n, 9H).

13B

(87) 17.8 mg, white solid. Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% DEA)/IPA; Gradient: 10% IPA; Detector: 254 nm], Retention time=12.34 min. LCMS (ES, m/z) [M+H]+: 522.40. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm) δ 7.13 (dd, J=4.2 Hz, J=8.7 Hz, 1H), 7.07 (dd, J=2.4 Hz, 6.9 Hz, 1H), 6.77 (td, J=2.4 Hz, 9.0 Hz, 1H), 6.58-6.53 (m, 2H), 5.19 (s, 1H), 4.58 (d, J=5.4 Hz, 1H), 4.42 (d, J=5.7 Hz, 1H), 4.01 (t, J=5.7 Hz, 2H), 3.71-3.65 (m, 1H), 3.56 (t, J=7.8 Hz, 2H), 3.24 (t, J=7.8 Hz, 2H), 3.06-2.99 (m, 1H), 2.91-2.86 (m, 3H), 2.57 (dd, J=2.7 Hz, J=14.7 Hz, 1H), 2.40 (dd, J=15.0 Hz, J=26.4 Hz, 1H), 1.23-1.10 (n, 9H).

(88) Using the similar procedures described above, the following additional compounds of the invention were prepared.

(89) TABLE-US-00002 TABLE 2 Exemplary Derivatives of Compound 13B Exam- ple No. Compound [M + H].sup.+ HNMR 14A 553.2 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 30% IPA; Detector: 254 nm], Retention time = 2.70 min. .sup.1HNMR (300 MHz, DMSO-d.sub.6, ppm): δ10.39 (s, 1 H), 7.14-7.10 (m, 3 H), 6.90 (d, J = 2.4 Hz, 1 H), 6.83-6.76 (m, 1 H), 5.47 (s, 1 H), 4.58 (d, J = 6.3 Hz, 1 H), 4.42 (d, J = 6.0 Hz, 1 H), 4.08-3.95 (m, 2 H), 3.72-3.68 (m, 1 H), 3.35-3.26 (m, 4 H), 3.04-2.91 (m, 3 H), 2.76- 2.56 (m, 3 H), 2.29-2.14 (m, 1 H), 1.16-1.09 (m, 9 H). 14B 553.2 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 30% IPA; Detector: 254 nm], Retention time = 3.12 min. .sup.1HNMR (300 MHz, DMSO-d.sub.6, ppm): δ10.40 (s, 1 H), 7.15-7.10 (m, 3 H), 6.90 (d, J = 2.4 Hz, 1 H), 6.83-6.76 (m, 1 H), 5.47 (s, 1 H), 4.58 (d, J = 6.6 Hz, 1 H), 4.42 (d, J = 6.0 Hz, 1 H), 4.05-3.93 (m, 2 H), 3.72-3.65(m, 1 H), 3.04 (s, 1 H), 3.35-3.26 (m, 4 H), 3.09-2.86 (m, 3 H), 2.73-2.56 (m, 3 H), 2.29-2.14 (m, 1 H), 1.16-1.04 (m, 9 H). 15A 534.3 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 30% EtOH; Detector: 254 nm], Retention time = 3.53 min. .sup.1HNMR (300 MHz, CDCl3, ppm) δ 7.38 (d, J = 9.3 Hz, 1H), 6.78-6.74 (m, 2H), 6.39 (d, J = 10.2 Hz, 2H), 5.15 (s, 1 H), 4.65-4.38 (m, 2H), 4.12-3.96 (m, 2H), 3.81 (s, 3H), 3.73-3.18 (m, 4H), 3.17-2.70 (m, 6 H), 2.63-2.23 (m, 2H), 1.28-1.04 (m, 9H). 15B 534.3 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 30% EtOH; Detector: 254 nm], Retention time = 4.77 min. .sup.1HNMR: (300 MHz, CDCl3, ppm): δ 7.38 (d, J = 9.3 Hz, 1H), 6.81-6.72 (m, 2H), 6.39 (d, J = 10.5 Hz, 2H), 5.15 (s, 1 H), 4.67- 4.39 (m, 2H), 4.25-3.95 (m, 2H), 3.81 (s, 3H), 3.73-3.22 (m, 4 H), 3.19-2.74 (m, 6 H), 2.59- 2.30 (m, 2H), 1.31-1.06 (m, 9H).

Example 3: Synthesis of Compound 16B

(90) ##STR00049## ##STR00050##

Step 1. Synthesis of 16-1

(91) To a solution of 3-bromo-1,1,1-trifluoropropan-2-one (10 g, 52.37 mmol) in trichloromethane (60 mL) at room temperature, was added a solution of NH.sub.2OH.HCl (5.46 g, 78.56 mmol) in water (10 mL) dropwise. The resulting solution was stirred at 65° C. for 24 h. After cooling to room temperature, the mixture was diluted with water (30 mL), and extracted with dichloromethane (40 mL×3). The organic phase was washed with brine (50 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by distillation under reduced pressure (0.1 MP) and the fraction was collected at 70-80° C. to afford the desired product (4.6 g, 43% yield) as a colorless oil.

Step 2. Synthesis of 16-2

(92) To a solution of (E)-N-(3-bromo-1,1,1-trifluoropropan-2-ylidene)hydroxylamine (4 g, 19.42 mmol,) in methyl-tert-butyl ether (800 mL), were added 1H-indole (9.1 g, 77.68 mmol) and sodium carbonate (12.5 g, 116.83 mmol). The resulting solution was then stirred at room temperature for 1 day. The solids were filtered out and the filter cake was washed with dichloromethane (50 mL×2). The filtrate was washed with brine (50 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum to afford the desired product (4 g, 85% yield) as light yellow oil.

Step 3. Synthesis of 16-3

(93) To a solution of 16-2 (4 g, 16.52 mmol) in tetrahydrofuran (100 mL) at 0° C., was added lithium aluminium hydride (2.6 g, 68.51 mmol) portion wise. The mixture was stirred at 0° C. for 10 min and then 70° C. overnight. The reaction mixture was cooled to room temperature and quenched by the addition of saturated aqueous NH.sub.4C.sub.1 (200 mL). The solid was filtered out and the filter cake was washed with ethyl acetate (100 mL×2). The filtrate was extracted with ethyl acetate (100 mL). The organic phase was washed with brine (200 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/3) as the eluent to afford the desired product (2.4 g, 64% yield).

Step 4. Synthesis of 16-4

(94) To a solution of 16-3 (264 mg, 1.16 mmol) and 2-fluoro-2-methylpropyl trifluoromethanesulfonate (430 mg, 1.92 mmol) in 1,4-dioxane (1 mL), were added 18-crown-6 (422 mg, 1.60 mmol) and N,N-diisopropylethylamine (620 mg, 4.80 mmol). The resulting solution was then stirred at 110° C. for 26 h. After cooling to room temperature, the reaction was concentrated under vacuum. The residue was dissolved in dichloromethane (50 mL), and washed with brine (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by Flash-Prep-HPLC with the following conditions [Column: C18 silica gel; Mobile phase: MeCN/H.sub.2O (0.1% TFA)), 25%-70% MeCN; Detector: 254 nm] to afford the desired product (270 mg).

Step 5. Synthesis of 16-5

(95) To a solution of 16-4 (600 mg, 1.98 mmol) in toluene (6 mL), were added 2,6-difluoro-4-iodobenzaldehyde (590 mg, 2.20 mmol) and acetic acid (0.2 mL). The reaction mixture was then stirred at 80° C. for 16 h. After cooled down, the mixture was concentrated under vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/29) as the eluent to afford the desired product (600 mg, 55% yield).

Step 6. Synthesis of 16-6

(96) To a solution of 16-5 (600 mg, 1.09 mmol) in ethane-1,2-diol (12 mL), were added 1,10-phenanthroline (20 mg, 0.11 mmol), CuI (105 mg, 0.55 mmol), and cesium carbonate (711 mg, 2.18 mmol). The reaction mixture was stirred at 100° C. for 2 h. After cooled down, the mixture was diluted with water (100 mL), and extracted with ethyl acetate (100 mL×2). The organic phase was washed with brine (100 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:2) as the eluent to afford the desired product (270 mg, 51% yield).

Step 7. Synthesis of 16-7

(97) To a solution of 16-6 (270 mg, 0.56 mmol) in tetrahydrofuran (10 mL), were added p-toluenesulfonyl chloride (128 mg, 0.67 mmol), 4-dimethylaminopyridine (13 mg, 0.11 mmol), and triethylamine (68 mg, 0.67 mmol). The reaction mixture was stirred at 30° C. for 16 h before concentrated under vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (2:1) as the eluent to afford the desired product (120 mg, 34% yield).

Step 8. Synthesis of 16

(98) To a solution of 3-(fluoromethyl)azetidine (261 mg, 1.39 mmol) in acetonitrile (10 mL), were added cesium carbonate (1.02 g, 3.13 mmol) and 16-7 (100 mg, 0.16 mmol). The reaction mixture was stirred at 80° C. for 16 h. After cooled down, the solids were filtered out and the filtrate was concentrated under vacuum. The residue was purified by Flash-Prep-HPLC [Column, C18 silica gel; Mobile phase (A: H.sub.2O (0.05% NH.sub.4HCO.sub.3), B:MeCN), 40% MeCN to 69% MeCN in 10 min; rate: 80 mL/min; Detector, UV 254 nm]. The crude product was then purified by Prep-HPLC [Column, Xbridge RP C18, 19×150 nm; Mobile phase (A: H.sub.2O (0.05% NH.sub.4HCO.sub.3), B: MeCN), MeCN=50% to MeCN=80% in 8 min, rate: 25 mL/min; Detector, UV 254 nm] to afford the desired product as racemate (25 mg, 29% yield).

(99) The racemic product 16 was resolved by Chiral-Prep-HPLC [Column, IA; Mobile phase, 12% ethanol/hexane (0.1% diethylamine); 20 mL/min; 18 min; Detector, 254 nm] to afford the desired products.

(100) Compound 16A: Retention time=7.7 min. LCMS (ES, m/z): 558.20 [M+H]+; .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.45 (d, J=6.9 Hz, 1H), 7.19 (d, J=6.9 Hz, 1H), 7.06-6.96 (m, 2H), 6.56 (d, J=11.1 Hz, 2H), 5.52 (s, 1H), 4.55 (d, J=5.4 Hz, 1H), 4.39 (d, J=5.4 Hz, 1H), 4.28-4.16 (m, 1H), 4.01-3.98 (m, 2H), 3.63-3.47 (m, 2H), 3.37-3.22 (m, 4H), 3.19-3.02 (m, 2H), 2.91-2.59 (m, 3H), 1.28-1.09 (m, 6H).

(101) Compound 16B: Retention time=14.5 min. LCMS (ES, m/z): 558.20 [M+H]+; .sup.1H-NMR (300 MHz, CD.sub.3OD, ppm): δ 7.44 (d, J=6.9 Hz, 1H), 7.20 (d, J=7.2 Hz, 1H), 7.05-6.96 (m, 2H), 6.56 (d, J=13.8 Hz, 2H), 5.52 (s, 1H), 4.55 (d, J=5.4 Hz, 1H), 4.40 (d, J=5.4 Hz, 1H), 4.28-4.02 (m, 1H), 4.01-3.98 (m, 2H), 3.58-3.53 (m, 2H), 3.34-3.25 (m, 4H), 3.13-3.10 (m, 2H), 2.92-2.88 (m, 2H), 2.73-2.59 (m, 1H), 1.28-1.09 (m, 6H).

Example 4: Synthesis of Compounds 17A and 17B and Derivatives Thereof

(102) ##STR00051## ##STR00052##

Step 1. Synthesis of 17-2

(103) To a solution of 1-(1-benzofuran-3-yl)propan-2-amine (1.2 g, 6.85 mmol) in 1,4-Dioxane (40 mL), were added N,N-diisopropylethylamine (2.6 g, 20.12 mmol) and 2-fluoro-2-methylpropyl trifluoromethanesulfonate (1.54 g, 6.87 mmol). The resulting solution was stirred at 75° C. for 16 h. After cooled down, the mixture was diluted with icy water (100 mL), and extracted with ethyl acetate (100 mL×3). The organic phase was washed with brine (100 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum to afford the desired product (1.5 g, 88% yield).

Step 2. Synthesis of 17-3

(104) To a mixture of 17-2 (820 mg, 3.29 mmol) in toluene (30 mL), were added acetic acid (593 mg, 9.88 mmol) and 2,6-difluoro-4-iodobenzaldehyde (883 mg, 3.29 mmol). The resulting solution was stirred at 100° C. for 2 days. After cooled down, the mixture was diluted with icy water (100 mL), and extracted with ethyl acetate (50 mL×3). The organic phase was washed with brine (50 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum to afford the desired product (0.3 g, 18% yield).

Step 3. Synthesis of 17-4

(105) To a mixture of 17-3 (300 mg, 0.60 mmol) in ethane-1,2-diol (10 mL), were added cesium carbonate (585 mg, 1.80 mmol) and copper(I) iodide (114.2 mg). The resulting solution was stirred at 100° C. overnight. After cooled down, the mixture was diluted with saturated ammonium chloride solution (30 mL), and extracted with ethyl acetate (30 mL×3). The organic phase was washed with brine (30 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum to afford the desired product (150 mg, 58% yield).

Step 4. Synthesis of 17-5

(106) To a solution of 17-4 (150 mg, 0.35 mmol) in dichloromethane (10 mL), were added triethylamine (35 mg, 0.35 mmol), 4-methylbenzene-1-sulfonyl chloride (79 mg, 0.41 mmol) and 4-dimethylaminopyridine (5 mg, 0.04 mmol). The resulting solution was stirred at room temperature overnight. The reaction was then quenched by water (30 mL). The resulting solution was extracted with dichloromethane (20 mL×3). The organic phase was washed with brine (20 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1:3) as the eluent to afford the desired product (0.17 g, 84% yield).

Step 5. Synthesis of 17A and 17B

(107) To a solution of 17-5 (170 mg, 0.29 mmol) in acetonitrile (10 mL) were added 3-(fluoromethyl)azetidine (103 mg, 1.16 mmol) and cesium carbonate (376 mg, 1.14 mmol). The resulting solution was stirred at 80° C. overnight. After cooled down, the mixture was diluted with icy water (30 mL), and extracted with ethyl acetate (30 mL×3). The organic phase was washed with brine (20 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by Prep-HPLC [Column: C18; Mobile Phase: CH.sub.3CN/H.sub.2O (0.05% NH.sub.4HCO.sub.3); Gradient: 70%-86% MeCN, 8 min; Detected: 254 nm] to afford the racemate 17 (30 mg, 21% yield) as a white solid.

(108) The racemate of 17 was resolved by Prep-Chiral-HPLC [Column: AD-H; Mobile Phase: Hex/EtOH; Gradient: 13% EtOH, 18 min; Rate: 20 mL/min; Detector: 254 nm] to afford the desired products.

(109) Compound 17A: Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 15% EtOH; Detector: 254 nm], Retention time=3.03 min. LC-MS (ES, m/z): 505.4 [M+H]+; .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.49-7.46 (m, 1H), 7.32-7.29 (m, 1H), 7.21-7.18 (m, 2H) 6.54 (d, J=10.8 Hz, 2H), 5.14 (s, 1H), 4.55 (d, J=5.7 Hz, 1H), 4.40 (d, J=5.4 Hz, 1H), 4.01-3.98 (m, 2H), 3.90 (d, J=5.4 Hz, 1H), 3.69-3.51 (m, 2H), 3.32-3.19 (m, 2H), 2.97-2.86 (m, 5H), 2.86-2.57 (m, 1H), 2.53-2.38 (m, 1H), 1.28-1.10 (min, 9H).

(110) Compound 17B: Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 15% EtOH; Detector: 254 nm], Retention time=5.20 min. LC-MS (ES, m/z): 505.4 [M+H]+; .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.47 (s, 1H), 7.30-7.19 (m, 3H), 6.54 (d, J=11.1 Hz, 2H), 5.14 (s, 1H), 4.55 (d, J=5.1 Hz, 1H), 4.40 (d, J=4.8 Hz, 1H), 4.06 (s, 2H), 3.68 (s, 1H), 3.56-3.51 (m, 2H), 3.31-3.20 (m, 2H), 3.07-2.88 (m, 5H), 2.67-2.33 (m, 2H), 1.63-1.40 (min, 9H).

(111) Using the similar procedures described above, the following additional compounds of the invention were prepared.

(112) TABLE-US-00003 TABLE 3 Exemplary Derivatives of Compound 17A and Compound 17B Exam- ple No. Compound [M + H].sup.+ HNMR 18 537.5 .sup.1H-NMR (300 MHz, CD.sub.3OD, ppm) δ 7.47- 7.44 (m, 1 H), 7.30-7.27 (m, 1 H), 7.19-7.17 (m, 2 H), 7.07 (s, 1H), 6.89 (s, 1 H), 5.59 (s, 1 H), 4.57 (d, J = 5.4 Hz, 1 H), 4.4 (d, J = 5.4 Hz, 1 H)), 4.03-4.00 (m, 2 H), 3.87-3.84 (m, 1 H), 3.56-3.51 (m, 2 H), 3.30-3.20 (m, 2 H), 3.11-3.07 (m, 1 H), 3.04-2.90(m, 1 H), 2.88- 2.78 (m, 3 H), 2.59 (d, J = 15.3 Hz, 1 H), 2.36-2.22 (m, 1 H), 1.37-0.90 (m, 9 H). 19A 509.4 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 70% IPA; Detector: 254 nm], Retention time = 12.27 min. .sup.1HNMR (300 MHz, CDCl3, ppm) δ 8.79 (s, 1H), 8.12 (d, J = 1.5 Hz, 1H), 7.81 (d, J = 7.5, 1H), 7.07-7.03 (m, 1H), 6.44 (d, J = 10.5 Hz, 2H), 5.25 (s, 1H), 4.60 (s, 1H), 4.44 (s, 1H), 4.26-3.90 (m, 4H), 3.68-3.58 (m, 3H), 3.24 (s, 2H), 3.09-3.00 (m, 2H), 2.67-2.57 (m, 2H), 1.45 (t, J = 18.6 Hz, 3H), 1.14 (d, J = 6.3 Hz, 3H). 19B 509.4 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 70% IPA; Detector: 254 nm], Retention time = 16.92 min. .sup.1HNMR (300 MHz, CDCl3, ppm) δ 8.86 (s, 1H), 8.12 (d, J = 1.5 Hz, 1H), 7.80 (d, J = 7.5 Hz, 1H), 7.06-7.02 (m, 1H), 6.43 (d, J = 10.5 Hz, 2H), 5.24 (s, 1H), 4.60 (s, 1H), 4.43 (d, J = 3.9 Hz, 1H), 4.26-3.95 (m, 4H), 3.63-3.59 (m, 3H), 3.24-3.00 (m, 4H), 2.72-2.17 (m, 2H), 1.44 (t, J = 18.6 Hz, 3H), 1.14 (d, J = 6.3 Hz, 3H).

Example 5: Synthesis of Compound 20

(113) ##STR00056##

Step 1. Synthesis of 20-1

(114) To a solution of 4-bromo-2-chloro-6-fluorobenzaldehyde (1.0 g, 4.21 mmol) in N,N-dimethylformamide (30 mL), were added triethylamine (840 mg, 8.30 mmol), Palladium acetate (47 mg, 0.21 mmol), P(o-tol).sub.3 (130 mg, 0.43 mmol), and ethyl prop-2-enoate (630 mg, 6.29 mmol). The reaction mixture was then purged three times with nitrogen and stirred at 100° C. overnight. After cooling to room temperature, the reaction mixture was then quenched by the addition of icy water. The mixture was extracted with dichloromethane (50 mL×3). The organic phase was washed with brine (30 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (1/5) as the eluent to afford the desired product (0.8 g, 74% yield).

Step 2. Synthesis of 20-2

(115) To a solution of (2R)-1-(1H-indol-3-yl)propan-2-amine (340 mg, 1.95 mmol) in toluene (10 mL), were added ethyl (2E)-3-(3-chloro-5-fluoro-4-formylphenyl)prop-2-enoate (600 mg, 2.34 mmol) and acetic acid (230 mg, 3.83 mmol). The reaction mixture was stirred at 80° C. overnight, then cooled to room temperature, and concentrated under vacuum. The residue was purified by silica gel column with ethyl acetate/petroleum ether (2/5) to afford the desired product (0.7 g, 87% yield).

Step 3. Synthesis of 20-3

(116) To a solution of ethyl 20-2 (700 mg, 1.70 mmol) in 1,4-dioxane (20 mL), were added N,N-diisopropylethylamine (1.09 g, 8.43 mmol) and 2-fluoro-2-methylpropyl trifluoromethanesulfonate (1.14 g, 5.09 mmol). The reaction mixture was stirred at 120° C. overnight, then cooled to room temperature, and concentrated under vacuum. The residue was purified by Prep-HPLC [Column: SunFire Prep C18, 5 um, 19*150 mm; Mobile Phase: MeCN/Water (0.1% FA); Gradient: 71%-86% MeCN, 6 min, 25 mL/min; Detector: 220 nm] to afford the desired product (0.25 g, 30% yield).

Step 4. Synthesis of 20

(117) To a solution of ethyl 20-3 (250 mg, 0.51 mmol) in tetrahydrofuran (30 mL) and water (3 mL), was added lithium hydroxide (25 mg, 1.04 mmol). The reaction mixture was stirred at room temperature overnight, and then concentrated under vacuum. The pH of the residue was adjusted to 5-6 with hydrogen chloride (1 N). The reaction was extracted with dichloromethane (30 mL×3). The organic phase was dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was dissolved in MeCN/H.sub.2O and dried by lyphilization to afford the desired product (75.4 mg, 32% yield). LCMS (ES, m/z): 459.16 [M+H]+; .sup.1HNMR: (300 MHz, DMSO-d.sub.6, ppm): δ 12.55 (br s, 1H), 10.44 (s, 1H), 7.70 (s, 1H), 7.59-7.52 (m, 2H), 7.41 (d, J=7.2 Hz, 1H), 7.17 (d, J=6.9 Hz, 1H), 7.01-6.92 (m, 2H), 6.69 (d, J=15.9 Hz, 1H), 5.35 (s, 1H), 3.68-3.61 (m, 1H), 3.08-2.90 (m, 2H), 2.64-2.59 (m, 1H), 2.50-2.20 (m, 1H), 1.16-1.05 (m, 9H).

Example 6: Synthesis of Compound 21

(118) ##STR00057##

Step 1. Synthesis of 21-1

(119) To a solution of (2R)-1-(1H-indol-3-yl)propan-2-amine (700 mg, 4.02 mmol) and 2-fluoro-2-methylpropyl trifluoromethanesulfonate (1.082 g, 4.80 mmol) in 1,4-dioxane (10 mL), was added N,N-diisopropylethylamine (1.56 g, 12.06 mmol). The resulting solution was stirred at 70° C. overnight. After cooling to room temperature, the mixture was diluted with water (50 mL), and extracted with ethyl acetate (100 mL×2). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to afford the desired product (830 mg, 83% yield).

Step 2. Synthesis of 21-2

(120) A solution of 21-1 (830 mg, 3.34 mmol), 4-bromo-2,6-dichlorobenzaldehyde (710 mg, 2.80 mmol) and acetic acid (350 mg, 5.83 mmol) in toluene (10 mL) was stirred at 80° C. overnight. The mixture was then cooled to room temperature, diluted with water (50 mL), and extracted with ethyl acetate (100 mL×2). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to afford the desired product (1 g, 74% yield).

Step 3. Synthesis of 21-3

(121) A solution of 21-2 (100 mg, 0.21 mmol), ethyl prop-2-enoate (31 mg, 0.31 mmol), Pd(OAc).sub.2 (95 mg, 0.42 mmol), Ph.sub.3P (110 mg), and triethylamine (42 mg, 0.42 mmol) in N,N-dimethylformamide (5 mL) was stirred at 100° C. for 48 h. After cooling to room temperature, the mixture was diluted with water (50 mL), and extracted with ethyl acetate (100 mL×2). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was dissolved in DMF and purified by Prep-HPLC with the following condition [Column: X Bridge Shield RP18 OBD, 5 um, 19×150 mm; Mobile phase A: Water (0.05% NH.sub.4HCO.sub.3), Mobile Phase B: MeCN; Gradient: 25% MeCN to 54% in 8 min; Detector: UV 254 nm] to afford the desired product (76 mg, 73% yield).

Step 4. Synthesis of 21

(122) To a solution of 21-3 (76 mg, 0.15 mmol) in tetrahydrofuran (5 mL) and water (1 mL) was added lithium hydroxide (13 mg, 0.31 mmol). The resulting solution was stirred at 25° C. for 15 h. The mixture was then diluted with water (20 mL). The pH of the solution was adjusted to 6 with hydrogen chloride (1 N), and extracted with ethyl acetate (50 mL×2). The organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was diluted with acetonitrile/water and dried by lyophilization to afford the desired product (15.6 mg, 22% yield). LCMS (ES, m/z): 476.38 [M+H].sup.+. .sup.1HNMR (300 MHz, CD.sub.3OD-d.sub.4, ppm): δ 7.66 (s, 1H), 7.44-7.34 (m, 3H), 7.17 (dd, J=1.8, 6.6 Hz, 1H), 7.00-6.92 (m, 2H), 6.54 (d, J=15.9 Hz, 1H), 5.71 (s, 1H), 3.87-3.83 (m, 1H), 3.22-3.19 (m, 1H), 3.16-2.97 (m, 1H), 2.68-2.64 (m, 1H), 2.37-2.22 (m, 1H), 1.14-1.11 (m, 6H), 1.09 (d, J=10.2 Hz, 3H).

Example 7: Synthesis of Compound 22

(123) ##STR00058##

Step 1. Synthesis of 22-2

(124) To a solution of 6-methoxy-1H-indole-3-carbaldehyde (2 g, 11.42 mmol) in toluene (20 mL), were added acetic acid (4.4 g, 57.08 mmol) and nitroethane (10 mL). The resulting solution was stirred at 130° C. for 6 h. After cooling to room temperature, the mixture was concentrated under vacuum. The resulting solution was diluted with ethyl acetate (100 mL). The mixture was washed with water (100 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to afford the desired product (2.5 g, crude).

Step 2. Synthesis of 22-3

(125) To a solution of 22-2 (2.5 g, 10.76 mmol) in tetrahydrofuran (100 mL) was added lithium aluminium hydride (1.63 g, 42.95 mmol) portion wise at 0° C. The resulting solution was then stirred at 65° C. for 4 h. After cooling to room temperature, the reaction was then quenched by the addition of icy water (100 mL), and extracted with ethyl acetate (100 mL×3). The organic phase was washed with water (100 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to afford the desired product (1.5 g, crude).

Step 3. Synthesis of 22-4

(126) A mixture of 1-(6-methoxy-1H-indol-3-yl)propan-2-amine (1.5 g, 7.34 mmol), 2-fluoro-2-methylpropyl trifluoromethanesulfonate (1.8 g, 8.03 mmol), and N,N-diisopropylethylamine (1.05 g, 8.08 mmol) in dioxane (20 mL) was stirred at 70° C. for 12 h. After cooling to room temperature, the mixture was diluted with water (50 mL), and extracted with ethyl acetate (50 mL×3). The organic phase was washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel column eluting with ethyl acetate/petroleum ether (1/10-1/1) to afford the desired product (1 g, 49% yield).

Step 4. Synthesis of 22-6A and 22-6B

(127) A mixture of (2-fluoro-2-methylpropyl)[1-(6-methoxy-1H-indol-3-yl)propan-2-yl]amine (1 g, 3.59 mmol), ethyl (2E)-3-(3,5-difluoro-4-formylphenyl)prop-2-enoate (860 mg, 3.58 mmol), and acetic acid (1 mL) in toluene (10 mL) was stirred at 80° C. for 12 h. After cooling to room temperature, the mixture was diluted with water (50 mL), and extracted with ethyl acetate (50 mL×3). The organic phase was concentrated under vacuum. The residue was purified by silica gel column eluting with ethyl acetate/petroleum ether (1/100-1/10) to afford the desired product as a racemate (750 mg). The racemate was separated by Chiral-Prep-HPLC [Column, IA; Mobile phase: Hex: EtOH=80:20, 20 mL/min; Detector: UV=254 nm] to afford 22-6A (RT=4.45 min) as a white solid and 22-6B (RT=9.27 min) as a white solid.

Step 5. Synthesis of 22A and 22B

(128) To a solution of 22-6A (30 mg, 0.06 mmol) in tetrahydrofuran (2 mL) and water (0.5 mL) was added lithium hydroxide (4.32 mg, 0.18 mmol). The resulting solution was stirred at room temperature for 12 h, then diluted with water (5 mL). The pH of the solution was adjusted to 6 with hydrogen chloride (1 N), and extracted with ethyl acetate (5 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated under vacuum to afford the desired product 22A (12.5 mg, 44% yield). LCMS (ES, m/z): 473.2 [M+H].sup.+. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ7.57 (d, J=16.2 Hz, 1H), 7.29 (m, 3H), 6.75 (d, J=2.4 Hz, 1H), 6.65 (dd, J=2.1, 8.4 Hz, 1H), 6.54 (d, J=15.9 Hz, 1H), 5.31 (s, 1H), 3.76 (s, 3H), 3.69-3.67 (m, 1H), 3.05-2.98 (m, 2H), 2.62-2.38 (m, 2H), 1.20-1.11 (m, 9H).

(129) In a similar manner, hydrolysis of 22-6B to obtain 22B. LCMS (ES, m/z): 473.2 [M+H].sup.+. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ7.57 (d, J=16.2 Hz, 1H), 7.29-7.18 (m, 3H), 6.75 (d, J=2.4 Hz, 1H), 6.65 (dd, J=2.1, 8.4 Hz, 1H), 6.54 (d, J=15.9 Hz, 1H), 5.31 (s, 1H), 3.76 (s, 3H), 3.69-3.67 (m, 1H), 3.05-2.98 (m, 2H), 2.62-2.38 (m, 2H), 1.20-1.11 (m, 9H).

Example 8: Synthesis of Compound 23

(130) ##STR00059## ##STR00060##

Step 1. Synthesis of 23-2

(131) To a solution of 5-(benzyloxy)-1H-indole-3-carbaldehyde (2 g, 7.96 mmol) in toluene (20 mL), were added nitroethane (24 mL) and ammonium acetate (600 mg, 8.00 mmol). The mixture was then stirred at 130° C. for 4 h. After cooling to room temperature, the mixture was diluted with water (100 mL), and extracted with ethyl acetate (50 mL×3). The organic phase was washed with brine (50 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum to afford the desired product (2.45 g, crude).

Step 2. Synthesis of 23-3

(132) To a solution of lithium aluminum hydride (1.2 g, 31.62 mmol) in tetrahydrofuran (30 mL), was added 5-(benzyloxy)-3-[(1Z)-2-nitroprop-1-en-1-yl]-1H-indole (2.45 g, 7.95 mmol) in tetrahydrofuran (20 mL) dropwise. The mixture was stirred at 0° C. for 30 min, and then stirred at 65° C. for 5 h. After cooling to room temperature, the mixture was diluted with icy water (100 mL), and extracted with ethyl acetate (100 mL×3). The organic phase was washed with brine (100 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford the desired product (2.2 g, crude).

Step 3. Synthesis of 23-4

(133) To a solution of 1-[5-(benzyloxy)-1H-indol-3-yl]propan-2-amine (1.2 g, 4.28 mmol) in 1,4-dioxane (30 mL), were added 2-fluoro-2-methylpropyl trifluoromethanesulfonate (960 mg, 4.28 mmol) and N,N-diisopropylethylamine (770 mg). The mixture was then stirred at 70° C. overnight. After cooling to room temperature, the mixture was diluted with water (100 mL), and extracted with ethyl acetate (50 mL×3). The organic phase was washed with brine (50 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (2/3) as the eluent to afford the desired product (430 mg, 28% yield).

Step 4. Synthesis of 23-5

(134) To a solution of [1-[5-(benzyloxy)-1H-indol-3-yl]propan-2-yl](2-fluoro-2-methylpropyl)amine (430 mg, 1.21 mmol) in methanol (20 mL) was added palladium on carbon (100 mg). The mixture was stirred at room temperature for 2 h under hydrogen atmosphere (1 atm). The mixture was filtered through Celite and the filtrate was concentrated under vacuum to afford the desired product (280 mg, 87% yield).

Step 5. Synthesis of 23-6

(135) To a solution of 3-[2-[(2-fluoro-2-methylpropyl)amino]propyl]-1H-indol-5-ol (280 mg, 1.06 mmol) in toluene (30 mL), were added ethyl (2E)-3-(3,5-difluoro-4-formylphenyl)prop-2-enoate (250 mg, 1.04 mmol) and acetic acid (200 mg, 3.33 mmol). The mixture was then stirred at 80° C. overnight. After cooling to room temperature, the mixture was diluted with water (100 mL), and extracted with ethyl acetate (50 mL×3). The organic phase was washed with brine (50 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was applied onto a silica gel column with ethyl acetate/petroleum ether (1:3) as the eluent to afford the desired product (180 mg, 36% yield).

Step 6. Synthesis of 23

(136) To a solution of 23-6 (100 mg, 0.21 mmol) in tetrahydrofuran (5 mL) and water (1 mL), was added lithium hydroxide (10 mg, 0.42 mmol). The mixture was then stirred at room temperature overnight, then diluted with water (20 mL). The pH of the solution was adjusted to 6 with hydrogen chloride (1 N), and extracted with ethyl acetate (20 mL×3). The organic phase was washed with brine (20 mL×2), dried over anhydrous sodium sulfate, and concentrated under vacuum to afford the desired product (11.9 mg, 13% yield). LCMS (ES, m/z): 459.18 [M+H]+; .sup.1HNMR: (300 MHz, CD.sub.3OD, ppm): δ7.53 (d, J=16.2 Hz, 1H), 7.18 (d, J=9.9 Hz, 2H), 6.99 (d, J=8.7 Hz, 1H), 6.81 (d, J=2.4 Hz, 1H), 6.59-6.50 (m, 2H), 5.26 (s, 1H), 3.69-3.63 (m, 1H), 3.08-2.87 (m, 2H), 2.56-2.50 (m, 1H), 2.46-2.32 (m, 1H), 1.32-1.09 (m, 9H).

Example 9: Synthesis of Compound 24 and Derivatives Thereof

(137) ##STR00061##

Step 1. Synthesis of 24-1

(138) To a solution of 1-fluorocyclopropane-1-carboxylic acid (700 mg, 6.73 mmol) in N,N-dimethylformamide (10 mL), were added N,N-diisopropylethylamine (1.5 g, 11.61 mmol), HATU (2.4 g, 6.32 mmol), and (2R)-1-(1H-indol-3-yl)propan-2-amine (1 g, 5.74 mmol). After stirring at room temperature overnight, the mixture was then quenched by the addition of icy water (20 mL). The mixture was extracted with ethyl acetate (20 mL×3). The organic phase was washed with brine (20 mL×3), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified on a silica gel column with ethyl acetate/petroleum ether (1:3) as the eluent to afford the desired product (600 mg, 40% yield).

Step 2. Synthesis of 24-2

(139) A solution of 24-1 (600 mg, 2.30 mmol) in BH.sub.3.THF (1 M) (20 mL) was heated to reflux overnight. After cooling to room temperature, the mixture was concentrated under vacuum, the residue was dissolved in methanol (20 mL), then heated to reflux overnight. After cooling to room temperature, the mixture was concentrated under vacuum. The residue was diluted with water (100 mL), and extracted with ethyl acetate (100 mL×3). The organic phase was washed with brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum. The crude mixture was purified on a silica gel column with ethyl acetate/petroleum ether (1:2) as the eluent to afford the desired product (350 mg, 62% yield).

Step 3. Synthesis of 24-3

(140) To a solution of (R)—N-((1-fluorocyclopropyl)methyl)-1-(1H-indol-3-yl)propan-2-amine (350 mg, 1.42 mmol) and ethyl (2E)-3-(3,5-difluoro-4-formylphenyl)prop-2-enoate (341.5 mg, 1.42 mmol) in toluene (5 mL), was added acetic acid (1 drop). After stirring at 80° C. for 2 h, the reaction mixture was cooled to room temperature, and concentrated under vacuum. The residue was purified on a silica gel column with ethyl acetate/petroleum ether (1:2) as the eluent to afford the desired product (120 mg, 18% yield).

Step 4. Synthesis of 24

(141) To a solution of 24-3 (120 mg, 0.26 mmol) in tetrahydrofuran (0.9 mL) and water (0.1 mL), was added lithium hydroxide (12.3 mg, 0.51 mmol). The resulting solution was stirred at room temperature overnight. The pH of the solution was adjusted to 6 with hydrogen chloride (1 N), and the mixture was extracted with ethyl acetate (20 mL×3). The organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified on a silica gel column with ethyl acetate/petroleum ether (1:1) as the eluent to afford the desired product (49.9 mg, 44% yield). LRMS (ES, m/z): 441.30[M+H].sup.+. .sup.1HNMR: (300 MHz, DMSO-d6, ppm): δ12.50 (br s, 1H), 10.59 (s, 1H), 7.56-7.40 (m, 4H), 7.19 (d, J=5.7 Hz, 1H), 700-7.6.95 (m, 2H), 6.67 (d, J=12 Hz, 1H), 5.23 (s, 1H), 3.58-3.56 (m, 1H), 3.09-2.88 (m, 2H), 2.67-2.63 (m, 2H), 1.07 (d, J=5.1 Hz, 3H) 0.94-0.89 (m, 2H), 0.56-0.51 (n, 2H).

(142) Using the similar procedures described above, the following additional compounds of the invention were prepared.

(143) TABLE-US-00004 TABLE 4 Exemplary Derivatives of Compound 24 Exam- ple No. Compound [M + H].sup.+ HNMR and Chiral HPLC 25 479.3 .sup.1HNMR (300 MHz, CDCl.sub.3, ppm) δ 7.65- 7.50 (m, 3 H), 7.42 (s, 1 H), 7.21 (s, 2 H), 7.15-7.08 (m, 2 H), 6.47 (d, J = 15.9 Hz, 1 H), 5.76 (s, 1 H), 3.76-3.69 (m, 1 H), 3.25- 3.07 (m, 2 H), 2.72 (d, J = 15.0 Hz, 1 H), 2.69-2.47 (m, 1 H), 1.45 (t, J = 18.9 Hz, 3 H), 1.16 (d, J = 6.6 Hz, 3 H). 26A 477.9 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 20% EtOH; Detector: 254 nm], Retention time = 3.70 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.47- 7.42 (m, 2 H), 7.26 (d, J = 11.1 Hz, 1 H), 7.12-7.03 (m, 2 H), 6.77-6.71 (m, 1 H), 6.54 (d, J = 8.1 Hz, 1 H), 5.44 (s, 1 H), 3.81-3.77 (m, 1 H), 3.16-3.10 (m, 1 H), 3.07-2.95 (m, 1 H), 2.59 (d, J = 14.7 Hz, 1 H), 2.40-2.25 (m, 1 H), 1.16 (s, 3 H), 1.11-1.09 (m, 6H) 26B 477.9 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 20% EtOH; Detector: 254 nm], Retention time = 3.17 min. .sup.1HNMR (300 MHz, CDCl.sub.3, ppm): δ 7.70- 7.58 (m, 1 H), 7.36 (s, 1 H), 7.17-7.08 (m, 3 H), 6.91-6.82 (m, 1 H), 6.43 (d, J = 16.5 Hz, 1 H), 5.43 (s, 1 H), 3.76-3.66 (m, 1 H), 3.21-3.16 (m, 1 H), 3.02-2.88 (m, 1 H), 2.59 (d, J = 14.4 Hz, 1 H), 2.35-2.14 (m, 1 H), 1.25 (s, 3 H), 1.19-1.09 (m, 6 H). 27A 493.3 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 10% IPA; Detector: 254 nm], Retention time = 4.19 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.64 (s, 1 H), 7.42 (s, 1 H), 7.37-7.32 (d, J = 15.6 Hz, 1 H), 7.12-7.02 (m, 2 H), 6.73 (dt, J = 2.4, 9.0 Hz, 1 H), 6.55 (d, J = 15.6 Hz, 1 H), 5.69(s, 1 H), 3.86-3.82 (m, 1 H), 3.18-2.96 (m, 2 H), 2.60 (d, J = 14.4 Hz, 1 H), 2.36-2.21 (m, 1 H), 1.20-1.07 (m, 9H). 27B 493.3 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 10% IPA; Detector: 254 nm], Retention time = 11.72 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.65 (s, 1 H), 7.43 (s, 1 H), 7.35 (d, J = 15.9 Hz, 1 H), 7.12-7.02 (m, 2 H), 6.73 (dt, J = 2.4, 9.0 Hz, 1 H), 6.55(d, J = 15.9 Hz, 1 H), 5.70(s, 1 H), 3.86-3.82 (m, 1 H), 3.17-2.96 (m, 2 H), 2.62 (d, J = 14.7 Hz, 1 H), 2.36- 2.21 (m, 1 H), 1.21-1.08 (m, 9 H). 28A 497.3 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 10% EtOH; Detector: 254 nm], Retention time = 5.21 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm) δ 7.80 (s, 1 H), 7.62-7.56 (m, 2H), 7.14 (dd, J = 4.2 Hz, J = 8.7 Hz, 1H), 7.09 (dd, J = 2.7 Hz, J = 9.9 Hz, 1H), 6.78 (td, J = 2.4 Hz, J = 9.3 Hz, 1H), 6.61 (d, J = 15.9 Hz, 1H), 5.81 (s, 1H), 3.79-3.71 (m, 1H), 3.22-3.14 (m, 2H), 2.72-2.67 (m, 1H), 2.64-2.53 (m, 1H), 1.43 (t, J = 18.9 Hz, 3H), 1.19 (d, J = 6.3 Hz, 3H). 28B 497.3 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 10% EtOH; Detector: 254 nm], Retention time = 10.04 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm) δ 7.80 (s, 1 H), 7.62-7.57 (m, 2H), 7.14 (dd, J = 4.5 Hz, J = 8.7 Hz, 1H), 7.09 (dd, J = 2.4 Hz, J = 9.9 Hz, 1H), 6.79 (td, J = 2.7 Hz, J = 9.3 Hz, 1H), 6.61 (d, J = 15.9 Hz, 1H), 5.81 (s, 1H), 3.79-3.71 (m, 1H), 3.22-3.10 (m, 2H), 2.72-2.67 (m, 1H), 2.65-2.54 (m, 1H), 1.43 (t, J = 18.6 Hz, 3H), 1.19 (d, J = 6.9 Hz, 3H). 29A 447.1 .sup.1H-NMR: (300 MHz, DMSO-d6, ppm): δ 12.57 (brs, 1 H), 10.61 (s, 1 H), 7.56-7.40 (m, 4 H),7.20 (d, J = 7.5 Hz, 1 H), 7.04- 6.93 (m, 2 H), 6.67 (d, J = 15.9 Hz, 1 H), 5.25 (s, 1 H), 3.47-3.41 (m, 1 H), 3.18- 3.04 (m, 1 H), 2.91-2.86 (m, 1 H), 2.64- 2.51 (m, 2H), 1.53-1.41 (m, 3 H), 1.10 (d, J = 6.3 Hz, 3 H). 30 0 437.5 .sup.1HNMR (300 MHz, DMSO-d.sub.6, ppm): 10.53 (s, 1 H), 7.55-7.39 (m, 4 H), 7.18 (d, J = 7.2 Hz, 1 H), 7.02-6.92 (m, 2 H), 6.67 (d, J = 15.9 Hz, 1 H), 5.12 (s, 1 H), 3.71-3.67 (m, 1 H), 2.97-2.91 (m, 1 H), 2.61-2.57 (m, 1 H), 2.30 (q, J = 12.9 Hz, 2 H), 0.98 (d, J = 6.3 Hz, 3 H), 0.91 (s, 3 H), 0.34-0.29 (m, 2 H), 0.19-0.11 (m, 2 H). 31A 497.4 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 20% EtOH; Detector: 254 nm], Retention time = 3.67 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.56 (d, J = 16.2 Hz, 1 H), 7.46 (d, J = 6.9 Hz, 1 H), 7.25-7.19 (m, 3 H), 7.07-6.92 (m, 2 H), 6.55 (d, J = 15.9 Hz, 1 H), 5.63 (s, 1 H), 4.22-4.18 (m, 1 H), 3.17-3.13 (m, 2 H), 2.74-2.6 (m, 2 H), 1.4-1.12 (m, 6 H). 31B 497.4 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 20% EtOH; Detector: 254 nm], Retention time = 4.03 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.49- 7.44(m, 2 H), 7.22-7.19 (m, 3 H), 7.06-6.97 (m, 2 H), 6.55(d, J = 15.9 Hz, 1 H), 5.62 (s, 1 H), 4.26-4.15 (m, 1 H), 3.17-3.08 (m, 2 H), 2.79-2.59 (m, 2 H), 1.44-1.12 (m, 6 H). 32A 444.5 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 20% EtOH; Detector: 254 nm], Retention time = 8.73 min. .sup.1HNMR (300 MHz, DMSO-d.sub.6, ppm): δ 8.11-8.09 (m, 1 H), 7.83 (dd, J = 1.5, 7.8 Hz, 1 H), 7.55 (d, J = 15.9 Hz, 1 H), 7.45 (d, J = 10.5 Hz, 2 H), 7.04-6.99 (m, 1 H), 6.67 (d, J = 15.9 Hz, 1 H), 5.25 (s, 1 H), 3.58-3.55 (m, 1 H), 2.93-2.80 (m, 2 H), 2.62-2.32 (m, 2 H), 1.26-1.05(m, 9 H). 32B 444.5 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 20% EtOH; Detector: 254 nm], Retention time = 11.3 min. .sup.1HNMR (300 MHz, DMSO-d.sub.6, ppm): δ 8.11-8.09 (m, 1 H), 7.84 (dd, J = 1.5, 7.8 Hz, 1 H), 7.53 (d, J = 16.2 Hz, 1 H), 7.43 (d, J = 10.5 Hz, 2 H), 7.04-6.99 (m, 1 H), 6.68 (d, J = 16.2 Hz, 1 H), 5.25 (s, 1 H), 3.58-3.55 (m, 1 H), 2.93-2.74 (m, 2 H), 2.62-2.28 (m, 2 H), 1.26-1.05 (m, 9 H). 33A 465.4 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 10% EtOH; Detector: 254 nm], Retention time = 9.26 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.58 (d, J = 15.9 Hz, 1 H), 7.23 (d, J = 10.2 Hz, 2 H), 7.15-7.06 (m, 2 H), 6.81-6.74 (m, 1 H), 6.54 (d, J = 15.9 Hz, 1 H), 5.33 (s, 1 H), 3.58-3.55 (m, 1 H), 3.20-2.95 (m, 2 H), 2.71-2.57 (m, 2 H), 1.50-1.37 (m, 3 H), 1.14 (d, J = 6.6 Hz, 3 H). 33B 465.4 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 10% EtOH; Detector: 254 nm], Retention time = 9.07 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm) δ 7.44 (d, J = 15.9 Hz, 1 H), 7.20-7.05 (m, 4 H), 6.81-6.74 (m, 1 H), 6.54 (d, J = 15.9 Hz, 1 H), 5.31 (s, 1 H), 3.58-3.55 (m, 1 H), 3.20- 2.95 (m, 2 H), 2.71-2.57 (m, 2 H), 1.50- 1.37 (m, 3 H), 1.14 (d, J = 6.6 Hz, 3 H). 34A 461.3 Chiral-Prep-HPLC [Column: IA, 100 mm, 4.6 mm, 20 mL/min, Mobile Phase: Hexane/Ethanol; Gradient: 4% EtOH; Rate: 20 mL/min; 20 min; Detector: 254 nm], Retention time = 13.64 min. .sup.1HNMR: (300 MHz, CD.sub.3OD, ppm): δ 7.56 (d, J = 13.8 Hz, 1 H), 7.35 (dd, J = 5.4, 8.4 Hz, 1 H), 7.20 (d, J = 9.9 Hz, 2 H), 6.87 (dd, J = 2.4, 9.9 Hz, 1 H), 6.78-6.71 (m, 1 H), 6.52 (d, J = 16.2 Hz 1 H), 5.27 (s, 1 H), 3.75-3.66 (m, 1 H), 3.06-2.88 (m, 2 H), 2.62-2.56 (m, 1 H), 2.47-2.33 (m, 1 H), 1.20-1.05 (m, 9 H). 34B 461.3 Chiral-Prep-HPLC [Column: IA, 100 mm, 4.6 mm, 20 mL/min, Mobile Phase: Hexane/Ethanol; Gradient: 4% EtOH; Rate: 20 mL/min; 20 min; Detector: 254 nm], Retention time = 18.38 min. .sup.1HNMR: (300 MHz, CD.sub.3OD, ppm): δ 7.56 (d, J = 13.8 Hz, 1 H), J = 13.8 Hz 1 H), 7.37-7.33 (dd, J = 5.1, 8.4 Hz, 1 H), 7.20 (d, J = 10.2 Hz, 2 H), 6.89-6.85 (dd, J = 2.4, 10.2 Hz, 1 H), 6.78-6.71 (m, 1 H), 6.52 (d, J = 16.2 Hz, 1 H), 5.27 (s, 1 H), 3.75- 3.65 (m, 1 H), 3.06-2.88 (m, 2 H), 2.62-2.56 (m, 1 H), 2.47-2.33 (m, 1 H), 1.23-1.09 (m, 9 H). 35A 465.3 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 10% IPA; Detector: 254 nm], Retention time = 7.59 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.57(d, J = 15.9 Hz 1 H), 7.36 (dd, J = 2.4, 8.4 Hz, 1 H), 7.23 (d, J = 9.9 Hz, 2 H), 6.89 (dd, J = 2.4, 9.9 Hz, 1 H), 6.79-6.72 (m, 1 H), 6.54 (d, J = 15.9 Hz, 1 H), 5.31 (s, 1 H), 3.58-3.54 (m, 1 H), 3.11-2.96 (m, 2 H), 2.67-2.57 (m, 2 H), 1.49-1.13 (m, 6 H). 35B 0 465.3 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 10% IPA; Detector: 254 nm], Retention time = 10.89 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.57(d, J = 15.9 Hz 1 H), 7.36(dd, J = 2.4, 8.4 Hz, 1 H), 7.23 (d, J = 9.9 Hz, 2 H), 6.89 (dd, J = 2.4, 9.9 Hz, 1 H), 6.79-6.72 (m, 1 H), 6.54 (d, J = 15.2 Hz, 1 H), 5.31 (s, 1 H), 3.58-3.54 (m, 1 H), 3.12-2.96 (m, 2 H), 2.67-2.57 (m, 2 H), 1.49-1.13 (m, 6 H). 36 459.3 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.38 (d, J = 15.6 Hz, 1 H), 7.21 (d, J = 8.4 Hz, 1 H), 7.11 (d, J = 10.2 Hz, 2 H), 6.62 (d, J = 2.1 Hz, 1 H), 6.56-6.49 (m, 2 H), 5.22 (s, 1 H), 3.66- 3.63 (m, 1 H), 3.07-2.92 (m, 2 H), 2.57-2.31 (m, 2 H), 1.21-1.08 (m, 9 H). 37A 473.2 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 20% IPA; Detector: 254 nm], Retention time = 7.37 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ7.53 (d, J = 15.9 Hz, 1 H), 7.18 (d, J = 10.2 Hz, 2H), 7.06 (d, J = 9 Hz, 1 H), 6.93 (d, J = 2.1 Hz, 1 H), 6.67 (dd, J = 2.4, 8.7 Hz, 1 H), 6.52(d, J = 15.9 Hz, 1 H), 5.34 (s, 1 H), 3.84 (s, 3 H), 3.69-3.67 (m, 1 H), 3.06-2.89 (m, 2 H), 2.62- 2.56 (m, 1 H), 2.47-2.03 (m, 1H), 1.32-1.00 (m, 9 H). 37B 473.2 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 20% IPA; Detector: 254 nm], Retention time = 5.54 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ7.53 (d, J = 15.9 Hz, 1 H), 7.18 (d, J = 9.9 Hz, 2 H), 7.06 (d, J = 8.4 Hz, 1 H), 6.93 (d, J = 2.1 Hz, 1 H), 6.67 (dd, J = 2.4, 8.7 Hz, 1 H), 6.52 (d, J = 15.9 Hz, 1 H), 5.27 (s, 1 H), 3.88 (s, 3 H), 3.81-3.66 (m, 1 H), 3.06-2.83 (m, 2 H), 2.61- 2.55 (m, 1 H), 2.46-2.33 (m, 1H), 1.32-0.90 (m, 9 H). 38A 477.9 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 20% EtOH; Detector: 254 nm], Retention time = 3.70 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.47- 7.42 (m, 2 H), 7.26 (d, J = 11.1 Hz, 1H), 7.12-7.03 (m, 2 H), 6.77-6.71 (m, 1 H), 6.54 (d, J = 8.1 Hz, 1 H), 5.44 (s, 1 H), 3.81-3.77 (m, 1 H), 3.16-3.10 (m, 1 H), 3.07-2.95 (m, 1 H), 2.59 (d, J = 14.7 Hz, 1 H), 2.40-2.25 (m, 1 H), 1.16 (s, 3 H), 1.11-1.09 (m, 6 H). 38B 477.9 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 20% EtOH; Detector: 254 nm], Retention time = 3.17 min. .sup.1HNMR (300 MHz, CDCl.sub.3, ppm): δ 7.70-7.58 (m, 1 H), 7.36 (s, 2 H), 7.17-7.08 (m, 3 H), 6.91-6.82 (m, 1 H), 6.43 (d, J = 16.5 Hz, 1 H), 5.43 (s, 1 H), 3.76-3.66 (m, 1 H), 3.21-3.16 (m, 1 H), 3.02-2.88 (m, 1 H), 2.59 (d, J = 14.4 Hz, 1 H), 2.35-2.14 (m, 1 H), 1.25 (s, 3 H), 1.19-1.09 (m, 6 H). 39A 443.5 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 15% EtOH; Detector: 254 nm], Retention time = 9.72 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.44 (d, J = 15.9 Hz, 1 H), 7.17-7.04 (m, 4 H), 6.79- 6.72 (m, 1 H), 6.53 (d, J = 15.9 Hz, 1 H), 5.18 (s, 1 H), 3.49-3.29 (m, 1 H), 2.94-2.92 (m, 1 H), 2.61-2.46 (m, 2 H), 2.21-2.08 (m, 1H), 1.72-1.69 (m, 1 H), 1.10 (d, J = 6.6 Hz, 3 H), 0.84 (d, J = 6.6 Hz, 3 H), 0.71 (d, J = 6.6 Hz, 3 H). 39B 443.5 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 15% EtOH; Detector: 254 nm], Retention time = 8.42 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.43 (d, J = 15.9 Hz, 1 H), 7.17-7.04 (m, 4 H), 6.79- 6.72 (m, 1 H), 6.53 (d, J = 15.9 Hz, 1 H), 5.18 (s, 1 H), 3.51-3.29 (m, 1 H), 2.98-2.92 (m, 1 H), 2.61-2.46 (m, 2 H), 2.21-2.08 (m, 1H), 1.74-1.69 (m, 1 H), 1.10 (d, J = 6.6 Hz, 3 H), 0.84 (d, J = 6.6 Hz, 3 H), 0.71 (d, J = 6.6 Hz, 3 H). 40A 457.4 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 30% EtOH; Detector: 254 nm], Retention time = 3.18 min. .sup.1H-NMR (300 MHz, CD.sub.3OD, ppm): δ 7.43- 7.34 (m, 2 H), 7.20-7.10 (m, 3 H), 7.03-6.94 (m, 2 H), 6.52 (d, J = 15.9 Hz, 1 H), 5.39 (s, 1 H), 3.31-3.24 (m, 1 H), 2.90-2.83 (m, 2 H), 2.75-2.65 (m, 1 H), 2.56-2.49 (m, 1 H), 1.75- 1.65 (m, 1 H), 1.45-1.35 (m, 1 H), 1.34-1.10 (m, 6 H), 0.91 (m, 3 H). 40B 457.4 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 30% EtOH; Detector: 254 nm], Retention time = 4.01 min. .sup.1H-NMR (300 MHz, CD.sub.3OD, ppm): δ 7.43- 7.34 (m, 2 H), 7.20-7.10 (m, 3 H), 7.03-6.94 (m, 2 H), 6.52 (d, J = 15.9 Hz, 1 H), 5.39 (s, 1 H), 3.31-3.24 (m, 1 H), 2.90-2.83 (m, 2 H), 2.75-2.65 (m, 1 H), 2.56-2.49 (m, 1 H), 1.75- 1.65 (m, 1 H), 1.45-1.35 (m, 1 H), 1.34-1.10 (m, 6 H), 0.91 (m, 3 H). 41 0 439.2 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.56 (d, J = 15.9 Hz, 1 H), 7.42-7.39 (m, 1 H), 7.25 (d, J = 11.1 Hz, 1 H), 7.18-7.12 (m, 2 H), 7.02- 6.93 (m, 2 H), 6.49 (d, J = 15.9 Hz, 1 H), 5.34 (s, 1 H), 3.86-3.82 (m, 1 H), 3.18-3.12 (m, 1 H), 3.02-2.93 (m, 1 H), 2.69 (d, J = 15 Hz, 1 H), 2.37-2.22 (m, 1 H), 2.01 (s, 3 H), 1.16-1.06 (m, 9 H). 42A 477.3 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 10% EtOH; Detector: 254 nm], Retention time = 9.58 min. .sup.1HNMR (300 MHz, CDCl.sub.3, ppm): δ 7.62 (d, J = 16.2 Hz, 1H), 7.53-7.42 (m, 2H), 7.19- 6.98 (m, 4H), 6.42 (d, J = 15.6 Hz, 1H), 5.32 (s, 1 H), 3.75-−3.60 (m, 1H), 3.14-2.71 (m, 2H), 2.67-2.25 (m, 2H), 1.39-0.99 (m, 9H). 42B 477.3 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 10% EtOH; Detector: 254 nm], Retention time = 7.11 min. .sup.1HNMR (300 MHz, CDCl.sub.3, ppm): δ 7.62 (d, J = 15.9 Hz, 1H), 7.52-7.42 (m, 2H), 7.18- 6.98 (m, 4H), 6.42 (d, J = 15.6 Hz, 1H), 5.31 (s, 1 H), 3.75-3.60 (m, 1H), 3.14-2.73 (m, 2H), 2.66-2.27 (m, 2H), 1.35-0.98 (m, 9H). 43A 477.2 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 30% IPA; Detector: 254 nm], Retention time = 3.15 min. .sup.1HNMR (300 MHz, CDCl.sub.3, ppm): δ 7.61 (d, J = 16.2 Hz, 1 H), 7.47 (s, 1 H), 7.42 (d, J = 8.4 Hz, 1 H), 7.21 (s, 1 H), 7.09-7.01 (m, 3 H), 6.42 (d, J = 15.9 Hz, 1 H), 5.30 (s, 1 H), 3.64-3.54 (m, 1 H), 3.08-3.01 (m, 1 H), 2.92- 2.80 (m, 1 H), 2.62-2.56 (m, 1 H), 2.47-2.34 (m, 1 H), 1.22 (t, J = 22.5 Hz, 6 H), 1.10 (d, J = 6.3 Hz, 3 H). 43B 477.2 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/IPA; Gradient: 30% IPA; Detector: 254 nm], Retention time = 2.72 min. .sup.1HNMR (300 MHz, CDCl.sub.3, ppm): δ 7.61 (d, J = 15.9 Hz, 1 H), 7.47 (s, 1 H), 7.42 (d, J = 8.1 Hz, 1 H), 7.21 (s, 1 H), 7.09-7.01 (m, 3 H), 6.42 (d, J = 15.9 Hz, 1 H), 5.30 (s, 1 H), 3.64-3.54 (m, 1 H), 3.08-3.01 (m, 1 H), 2.92- 2.80 (m, 1 H), 2.62-2.56 (m, 1 H), 2.47-2.34 (m, 1 H), 1.22 (t, J = 22.2 Hz, 6 H), 1.11 (d, J = 6.6 Hz, 3 H). 44A 448.5 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 15% EtOH; Detector: 254 nm], Retention time = 4.48 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.57 (d, J = 15.9 Hz, 1 H), 7.55-7.51 (m, 1 H), 7.50- 7.49 (m, 1 H), 7.35-7.32 (m, 1 H), 7.30-7.19 (m, 3 H), 6.56 (d, J = 16.2 Hz, 1 H), 5.29 (s, 1 H), 3.62-3.56 (m, 1 H), 3.15-3.14 (m, 1 H), 3.11-2.95 (m, 1 H), 2.69-2.60 (m, 2 H), 1.44 (t, J = 18.6 Hz, 3 H), 1.18 (d, J = 6.6 Hz, 3 H). 44B 448.5 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 15% EtOH; Detector: 254 nm], Retention time = 3.69 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.57 (d, J = 15.9 Hz, 1 H), 7.55-7.51 (m, 1 H), 7.50- 7.49 (m, 1 H), 7.35-7.32 (m, 1 H), 7.30-7.19 (m, 3 H), 6.56 (d, J = 16.2 Hz, 1 H), 5.29 (s, 1 H), 3.62-3.56 (m, 1 H), 3.15-3.14 (m, 1 H), 3.11-2.95 (m, 1 H), 2.69-2.60 (m, 2 H), 1.44 (t, J = 18.6 Hz, 3 H), 1.18 (d, J = 6.6 Hz, 3 H). 45 476.4 .sup.1HNMR (300 MHz, CD.sub.3OD, ppm) δ 7.75- 7.65 (m, 1 H), 7.49-7.41 (m, 2 H), 7.39-7.31 (m, 1 H), 7.29-7.22 (m, 1 H), 7.20-7.15 (m, 2 H), 6.56 (d, J = 15.6 Hz, 1 H), 5.69 (s, 1H), 3.90-3.86 (m, 1 H), 3.16-2.99 (m, 2 H), 2.62 (d, J = 15.3 Hz, 1 H), 2.36-2.21 (m, 1H), 1.24-1.09 (m, 9 H). 46A 461.3 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 10% EtOH; Detector: 254 nm], Retention time = 9.39 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm) δ 7.58 (d, J = 15.9 Hz, 1 H), 7.20 (d, J = 9.9 Hz, 2 H), 7.12-7.01 (m, 2 H), 6.79 (dt, J = 2.7, 9.3 Hz, 1 H), 6.53 (d, J = 16.2 Hz, 1 H), 5.29 (s, 1 H), 3.70-3.65 (m, 1 H), 3.05-2.88 (m, 2 H), 2.65- 2.55 (m, 1 H), 2.47-2.34 (m, 1 H), 1.29-1.09 (m, 9 H). 46B 461.3 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 10% EtOH; Detector: 254 nm], Retention time = 8.20 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm) 7.57 (d, J = 23.7 Hz, 1 H), 7.20-7.05 (m, 4 H), 6.76 (dt, J = 2.1, 9.3, 1 H), 6.53 (d, J = 16.2 Hz, 1 H), 5.28 (s, 1 H), 3.69-3.67 (m, 1 H), 3.05- 2.88 (m, 2 H), 2.60-2.33 (m, 2 H), 1.40-1.10 (m, 9 H). 47A 00 444.4 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 5% EtOH; Detector: 254 nm], Retention time = 5.58 min. .sup.1H NMR (300 MHz, DMSO-d.sub.6, ppm): δ7.59 (d, J = 4.2 Hz, 1 H), 7.57-7.44 (m, 4 H), 7.28- 7.23 (m, 2 H), 6.68 (d, J = 15.9 Hz, 1 H), 5.21 (s, 1 H), 3.51-343 (m, 1 H), 2.98-2.85 (m, 2 H), 2.62-2.57 (m, 1 H), 2.51-2.44 (m, 1 H), 1.29-1.09 (m, 9 H). 47B 01 444.4 Chiral HPLC [Column: IA, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 5% EtOH; Detector: 254 nm], Retention time = 6.17 min. .sup.1H NMR (300 MHz, DMSO-d.sub.6, ppm): δ7.58- 7.43 (m, 5 H), 7.28-7.21 (m, 2 H), 6.68 (d, J = 15.9 Hz, 1 H), 5.21 (s, 1 H), 3.53-3.51 (m, 1 H), 2.98-2.85 (m, 2 H), 2.62-2.57 (m, 1 H), 2.52-2.36 (m, 1 H), 1.25-1.09 (m, 9 H). 48A 02 460.3 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 30% EtOH; Detector: 254 nm], Retention time = 3.37 min. .sup.1HNMR: (300 MHz, CD.sub.3OD, ppm): δ 7.72 (d, J = 7.8 Hz, 1 H), 7.66 (d, J = 7.8 Hz, 1 H), 7.39-7.24 (m, 3 H), 7.13 (d, J = 10.2 Hz, 2 H), 6.53 (d, J = 15.9 Hz, 1 H), 5.34 (s, 1 H), 3.80- 3.78 (m, 1 H), 3.20-2.91 (m, 2 H), 2.78-2.72 (m, 1 H), 2.45-2.31 (m, 1 H), 1.19-1.11 (m, 9 H). 48B 03 460.3 Chiral HPLC [Column: AD, 100 mm, 4.6 mm, 0.6 mL/min, Mobile Phase: hexane (0.1% TFA)/ethanol; Gradient: 30% EtOH; Detector: 254 nm], Retention time = 2.66 min. .sup.1HNMR (300 MHz, CD.sub.3OD, ppm): δ 7.72 (d, J = 7.8 Hz, 1 H), 7.66 (d, J = 7.8 Hz, 1 H), 7.38-7.25 (m, 3 H), 7.12 (d, J = 10.2 Hz, 2 H), 6.53 (d, J = 15.9 Hz, 1 H), 5.34 (s, 1 H), 3.81- 3.77 (m, 1 H), 3.20-2.91 (m, 2 H), 2.78-2.72 (m, 1 H), 2.45-2.31 (m, 1 H), 1.19-1.11 (m, 9 H). 49 04 463.1 .sup.1H NMR (300 MHz, DMSO-d.sub.6, ppm): δ 12.59 (s, 1H), 10.52 (s, 1H), 7.73 (s, 1H), 7.67- 7.51 (m, 2H), 7.42 (dd, J = 7.1, 1.6 Hz, 1H), 7.23-7.13 (m, 1H), 7.03-6.93 (m, 2H), 6.71 (d, J = 16.0 Hz, 1H), 5.42 (s, 1H), 3.67-3.48 (m, 1H), 3.23-2.95 (m, 2H), 2.72-2.53 (m, 2H), 1.39 (t, J = 19.1 Hz, 3H), 1.09 (d, J = 6.3 Hz, 3H).

Biological Assays of the Exemplary Compounds Described Herein

Example 9. MCF-7 ER Degradation Assay

(144) The level of ER degradation by SERD described in this patent was analyzed by cell based high content imaging method. Briefly, human ER+ breast cancer MCF-7 cells were seeded into Grenier 96-well plates at a density of 5000 cells per well and grown in RPMI 1640 supplemented with 10% FBS (Gibco) at 37° C. with 5% CO.sub.2 for 48 hours. Then, 1 μL of compound solution (in DMSO) was added to the cells to make a final concentration of 1 nM per each well of treated cells. DMSO at the same final concentration (%) per each well was used to serve as negative control on each plate. After 4 hours or 24 hours of treatment at 37° C. with 5% CO.sub.2, the cells were fixed and permeabilized. Media from each well was aspirated and cells were gently washed with 1×PBS for 5 minutes and the wash procedure was repeated 3 times. Cells was then fixed with 4% formaldehyde freshly made in 1×PBS at room temperature for 20 minutes, followed by gentle washing with 1×PBS, 3 times for 5 minute each. The assay plate was incubated with 200 μl of 1×PBS containing 0.2% Triton-X100 for 5 minutes at room temperature, cells were then washed again with 1×PBS 3 times for 5 minutes each. After this, cells were incubated with the blocking buffer (1×PBS with 5% BSA) at room temperature for 1 hour. For immunostaining, blocking buffer was removed and primary anti-ER antibody (ER1D5, Santa Cruz) diluted in blocking buffer (1:400) was added to each well and cells were incubated overnight at 4° C. The next day, cells were washed with 1×PBS for 3 times with 5 minute each. Fluorochrome-labeled secondary antibody (goat anti-mouse IgG antibody, Alexa Fluor 488 conjugate, ThermoFisher, Cat #: A-11001) diluted in 1% BSA in 1×PBS (1:1000) was added to each well and incubated with the cells for 45 min at 37° C. in a moist environment in the dark. Cells were washed with 1×PBS for 3 times with 5 minute each time. Cells were incubated with DAPI (Beyotime; 1 g/ml) for 10 minutes and washed with 1×PBS for 3 times with 5 minute each time. For high content imaging analysis, plates were read on Cellomics ArrayScan™ XTI High Content Platform (ASN00002P) with excitation at 485 nm and imaging data was collected with minimum of 1000 cells per well. SERD induced ER degradation in the MCF-7 cells were measured using DMSO negative control as the baseline.

(145) TABLE-US-00005 TABLE 6 ER Degradation in MCF-7 cells MCF7 ER MCF7 ER Degradation at 4 hr Degradation at 24 hr Compound (%) (%) Fulvestrant 29 15 AZD9496 17 23 20 25 21 34A 21 20 46A 25 20

Example 10. MCF-7 Cell Growth Study

(146) Compound activity was analyzed using celltiter-glo luminescent viability assay (Promega #G7572) in breast cancer cell line MCF-7. Cells grown in log phase are trypsinized and seeded into a 96-well cell culture plate at a density of 2×10.sup.3 per well and incubated overnight at 37° C. with 5% CO.sub.2 in humidified culture chamber. The next day, compounds were prepared in 100% DMSO and were serially diluted and added into cells in the following final concentrations: 316, 100, 31.6, 10, 3.16, 1, 0.32, 0.1, 0.03, 0.01, 0.003 and 0.001 nM. As a control, same volume and concentration of DMSO vehicle control solution was added to the control wells in each plate. The cells were incubated with compounds at 37° C. culture chamber for 6 days. Promega's celltiter-glo kit was used to analyze the cell viability, according to manufacture's instruction. Briefly, fifty microliter of celltiter-glo reagent was added to each well, plate (covered with aluminum foil) was gently vibrated for 10 min to induce cell lysis at room temperature. Luminescence was measured using a SPECTRAmax i3 reader. Cell growth inhibition IC.sub.50 is calculated using GraphPad Prism V5.0 software. % inhibition=(1−(max signal/min signal))*100%

(147) TABLE-US-00006 TABLE 5 Inhibitory Activity of Representative Compounds Compound IC.sub.50 (nM) Compound IC.sub.50 (nM) Fulvestrant 0.4 AZD9496 0.4 GDC-0810 8.5  1 0.3  2 0.9  3 0.6  4 0.9  5 0.3  6 0.4  7 0.8  8 1  9 3.1 10 3.2 11 0.4 12 0.26 13A 0.5 14B 2.6 15A 1.8 16A 0.6 17A 0.8 18 8.9 19A 3.1 20 0.3 21 0.5 22A 2.6 23 4.2 24 1.9 25 0.8 26A 0.6 27B 0.7 28 1.3 29 0.9 30 1 31A 0.4 32A 1.9 33A 1.2 34A 0.4 35A 2.5 36 16 37A 2.9 38A 0.6 39A 2.6 40A 0.9 40B >100 41 0.6 42A 1.9 42B >100 43B 0.8 44A 7.9 45 8.9 46A 0.4 46B >100 47A >100 47B 0.6 48A 4.7 49 0.5

Example 11. Human Hepatocyte Clearance Study

(148) The in vitro hepatocyte clearance of compound described here was studied using pooled human hepatocytes purchased from BioreclamationIVT (Westbury, N.Y., Cat #X008001, Lot #TQJ). Assay was conducted according to manufacture's instruction. Briefly, 10 mM tock solutions of test compound and positive control (Verapamil) were prepared in 100% DMSO. Thawing media (50 mL) used in the study consists of: 31 mL Williams E medium (GIBCO Cat #12551-032); 15 mL isotonic percoll (GE Healthcare Cat #17-0891-09); 500 uL 100×GlutaMax (GIBCO Cat #35050); 750 uL HEPES (GIBCO Cat #15630-080); 2.5 mL FBS (Corning Cat #35-076-CVR); 50 uL human insulin (GIBCO Cat #12585-014) and 5 uL dexamethasone (NICPBP). Incubation media is made of Williams E medium supplemented with 1×GlutaMax. Thawing medium and supplement incubation medium (serum-free) were placed in a 37° C. water bath for at least 15 minutes prior to use. Compound stock solutions were diluted to 100 μM by combining 198 μL of 50% acetonitrile/50% water and 2 μL of 10 mM stock solution. Verapamil was use as positive control in the assay. Vials of cryopreserved hepatocytes were removed from storage and thawed in a 37° C. water bath with gentle shaking. Contents of the vial were poured into the 50 mL thawing medium conical tube. Vials were centrifuged at 100 g for 10 minutes at room temperature. Thawing medium was aspirated and hepatocytes were re-suspended with serum-free incubation medium to yield ˜1.5×10.sup.6 cells/mL. Hepatocyte viability and density were counted using a Trypan Blue exclusion, and then cells were diluted with serum-free incubation medium to a working cell density of 0.5×10.sup.6 viable cells/mL. Then, a portion of the hepatocytes at 0.5×10.sup.6 viable cells/mL was boiled for 5 min prior to adding to the plate as negative control to eliminate the enzymatic activity so that little or no substrate turnover should be observed. The boiled hepatocytes were used to prepare negative samples. Aliquots of 198 μL hepatocytes were dispensed into each well of a 96-well non-coated plate. The plate was placed in the incubator on an orbital shaker at 500 rpm for approximately 10 minutes. Aliquots of 2 μL of the 100 μM test compound or positive control were added into respective wells of the non-coated 96-well plate to start the reaction. This assay was performed in duplicate. The plate was incubated in the incubator on an orbital shaker at 500 rpm for the designed time points. Twenty-five microliter of contents were transferred and mixed with 6 volumes (150 μL) of cold acetonitrile with IS (200 nM imipramine, 200 nM labetalol and 200 nM diclofenac) to terminate the reaction at time points of 0, 15, 30, 60, 90 and 120 minutes. Samples were centrifuged at 3,220 g for 25 minutes and aliquots of 150 μL of the supernatants was used for LC-MS/MS analysis. For data analysis, all calculations were carried out using Microsoft Excel. Peak areas were determined from extracted ion chromatograms. The in vitro half-life (t.sub.1/2) of parent compound was determined by regression analysis of the percent parent disappearance vs. time curve. The in vitro half-life (in vitro t.sub.1/2) was determined from the slope value: in vitro t.sub.1/2=0.693/k. Conversion of the in vitro t.sub.1/2 (in min) into the scale-up unbound intrinsic clearance (Scaled-up unbound CL.sub.int, in mL/min/kg) was done using the following equation (mean of duplicate determinations): Scaled-up unbound CL.sub.int=kV/N×scaling factor, where V=incubation volume (0.5 mL); N=number of hepatocytes per well (0.25×10.sup.6 cells). Scaling factors for in vivo intrinsic clearance prediction using human hepatocytes are listed as: liver weight (g liver/kg body weight): 25.7; hepatocyte concentration (10.sup.6 cells/g liver): 99; scaling factor: 2544.3.

(149) TABLE-US-00007 TABLE 7 Human Hepatocyte Clearance of Selected SERD Human Hepatocyte Human In Human Remaining Human In vitro Cl.sub.int Scale-up Percentage @ vitro T.sub.1/2 (μL/min/ Cl.sub.int Compound 120 min (%) (min) 10.sup.6 cells) (mL/min/kg) Fulvestrant 12 42 33 86 GDC-0810 24 56 24.6 63 AZD9496 58 153 9.0 23  1 88 853 1.6 4.1 11 83 664 2.0 5.3 20 70 246 5.6 14 21 84 586 2.4 6 29 83 571 2.4 6.2

Example 12. Mouse PK Studies

(150) Mouse PK study (iv 3 mpk, and po 30 mpk) was conducted using male CD1 mice (25-33 g) obtained from SLAC Laboratory Animal Co., LTD of Shanghai). Compound is prepared with the following formulation: 5% DMSO, 5% Solutol HS 15 and 9% HPBCD in water. For intravenous (iv) dose at 3 mpk and oral (po) dose at 30 mpk, the compound is formulated at the concentration 0.2 mg/mL and 1 mg/mL, respectively. Compound formulation is freshly made prior to dosing in the morning. Oral dosing is via the use of oral gavage at 10 mL/kg, while iv dosing is via tail vein at 5 mL/kg. Three mice are used for each dosing route per compound per dose. Serial blood sample (30 uL whole blood at each time point) is collected into the K.sub.2EDTA tubes via facial vein at the following time points: 5 min (for iv route only), 30 min, 1, 2, 4, 6, 8, 12 and 24 hours. For each blood sample collected, immediately transfer 20 uL blood sample and mix well with 60 uL water in a 96-well plate (whole blood:water=1:3 v/v). Diluted blood samples are stored at −80 C freezer until analysis. For analysis, an aliquot of 20 μL sample was added with 200 μL IS (Glipizide, 100 ng/mL) in ACN. The mixture was vortexed for 2 min and centrifuged at 5800 rpm for 10 min. An aliquot of 2 μL supernatant was injected for LC-MS/MS analysis. Compound at 2.0-3000 ng/mL in CD1 mouse diluted blood is used for calibration curve. Pharmacokinetic data are obtained and described in the table below:

(151) TABLE-US-00008 TABLE 8 PK Data of Selected SERD Compounds Mouse Mouse IV 3 mpk Mouse PO PO 30 mpk AUC Compound CL (L/hr/kg) 30 mpk T½ (hr) last (uM .Math. hr) GDC-0810 0.92 2.9 80 AZD9496 1.14 3.5 85 21 0.11 9.2 412 25 0.08 62 658

Example 13. Human Breast Cancer xMCF-7 Xenograft Efficacy Study in Mouse

(152) To investigate the in vivo efficacy of SERD compounds described in this application, female nude mice were inoculated with human ER+ breast cancer cells xMCF-7. xMCF-7 cells are derived from MCF-7 (ATCC) tumor grown in the nude mice. Briefly, female balb/c mice (age 6-7 wk) were first inoculated subcutaneously on the back with estrogen pellet (0.5 mg, 60-day release from Innovative Research of America, Cat #SE-121). Two days later, each mouse was inoculated with 5 million xMCF-7 cells, prepared as 0.2 ml cell suspension in 1:1 mix of Eagle's MEM to cell culture media and Matrigel (Corning Cat #354234) for each injection. After tumor size reached −235 mm.sup.3, mice with xMCF-7 xenograft tumor are randomized to 10 mice per group and started receiving drug treatment. Each compound was prepared in dosing vehicle (5% DMSO, 5% Solutol HS15 and 10% HPBCD in water) and given orally at 5 mg/kg or 30 mg/kg, once a day for 28 days, except fulvestrant was prepared in peanut oil and given at 250 mg/kg subcutaneously, once a day for 28 days. In this study, drug treatment resulted in almost complete tumor growth inhibition, with compound 25 shown to be more efficacious in vivo than fulvestrant, GDC-0810 and AZD9496 (FIG. 1).

(153) In another xMCF-7 study, treatment initiated when tumor reached −250 mm.sup.3. Each drug was given at 2 mg/kg orally, once daily for 28 days. Significant tumor growth inhibition and tumor regression was observed and compounds 12, 21 and 25 were shown to be more efficacious than AZD9496 (FIG. 2).

Example 14: Combination of SERD Compound 25 with CDK4/6 Inhibitor Palbocilib in xMCF-7 Xenograft Model

(154) CDK4/6 inhibitor such as pabociclib was approved for ER positive, Her2 negative metastatic breast cancer, to investigate if combination of SERD and CDK4 inhibitor would further increase the efficacy in ER positive breast cancer, we tested SERD compound 25 in combination with CDK4/6 inhibitor palbociclib in xMCF-7 human breast cancer xenograft model. In this study, when xMCF-7 tumor reached −250 mm.sup.3, compound 25 was given orally, once daily for 28 days and palbociclib was given orally at 50 mg/kg, once daily for 14 day. As shown in FIG. 3, combination of palbociclib and compound 25 results in better tumor growth regression than either palbociclib or compound 25 used alone.

EQUIVALENTS AND SCOPE

(155) In the claims, articles such as “a,” “an,” and “the” may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include “or” between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The invention includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The invention includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process.

(156) Furthermore, the invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the invention, or aspects of the invention, is/are referred to as comprising particular elements and/or features, certain embodiments of the invention or aspects of the invention consist, or consist essentially of, such elements and/or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms “comprising” and “containing” are intended to be open and permits the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

(157) This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present invention that falls within the prior art may be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the invention can be excluded from any claim, for any reason, whether or not related to the existence of prior art.

(158) Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description, but rather is as set forth in the appended claims. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present invention, as defined in the following claims.