WO2019118855A1

WO2019118855A1 - Compounds and compositions for prevention and/or treatment of cancer

Info

Publication number: WO2019118855A1
Application number: PCT/US2018/065708
Authority: WO
Inventors: Todd RIDKY; Christopher Natale
Original assignee: University of Pennsylvania Penn
Current assignee: University of Pennsylvania Penn
Priority date: 2017-12-14
Filing date: 2018-12-14
Publication date: 2019-06-20
Anticipated expiration: 2020-06-14

Abstract

The present invention includes compounds, compositions and methods that are useful for preventing or treating certain types of cancer in a subject. In certain embodiments, the compounds comprise estrogen (including estrogen derivatives or analogues) and/or a selective GPER agonist.

Description

Compounds and Compositions for Prevention and/or Treatment of Cancer

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Application No. 62/598,776, filed December 14, 2017, which-application is incorporated herein by reference in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR

DEVELOPMENT

This invention was made with government support under grant number

F31CA206325 awarded by the National Cancer Institute. The government has certain rights in the invention.

BACKGROUND OF THE INVENTION

Cancer is the term used to describe a group of diseases involving abnormal cell growth with the potential to invade or spread to other parts of the body. Over 100 types of cancers affect humans. All tumor cells show six hallmarks, which are required to produce a malignant tumor: cell growth and division absent proper signals; continuous growth and division even given contrary signals; avoidance of programmed cell death; limitless number of cell divisions; blood vessel generation; and tissue invasion and metastases formation.

Cancer is characterized by failed tissue growth regulation. For a normal cell to transform into a cancer cell (a phenomenon described as malignant transformation), genes that regulate cell growth and differentiation must be altered. Such genes are divided into two broad categories: oncogenes (which promote cell growth and reproduction), and tumor suppressor genes (which inhibit cell division and survival). Malignant transformation can occur through formation of novel oncogenes, inappropriate over-expression of normal oncogenes, or under-expression or disabling of tumor suppressor genes. Typically, malignant transformations require changes in multiple genes: through large-scale mutations (including transposition, deletion, or gain of a chromosome or part thereof), or small-scale mutation (including point mutations, deletions, and insertions of a specific gene; or integration of genomic material from a DNA virus or retrovirus).

There is a need in the art for compounds, compositions, and methods that can be used to prevent and/or treat cancer in a subject. Such compounds, compositions, and methods should exhibit equivalent and/or superior clinical efficacy to current anticancer therapeutics, or alternatively, increase the efficacy of other anticancer therapeutics when used in combination therapy. The present invention satisfies this need.

BRIEF SUMMARY OF THE INVENTION

The invention provides a method of treating or preventing pancreatic cancer in a subject. The invention further provides a method of treating or preventing Merkel cell carcinoma (MCC) in a subject. The invention further provides a composition comprising (i) estrogen and/or GPER agonist and (ii) an immune checkpoint inhibitor. The invention further provides a kit comprising (i) estrogen and/or GPER agonist and (ii) an immune checkpoint inhibitor, and instructional material for use thereof in treating or preventing a cancer in a subject.

In certain embodiments, the method comprises administering to the subject in need thereof a therapeutically effective amount of estrogen and/or a selective G protein-coupled estrogen receptor (GPER) agonist. In certain embodiments, the method comprises administering to the subject in need thereof a therapeutically effective amount of estrogen and/or a GPER agonist.

In certain embodiments, the subject is further administered at least one additional anticancer agent. In certain embodiments, the subject is further administered at least one immunotherapeutic agent.

In certain embodiments, the pancreatic cancer is K-Ras driven.

In certain embodiments, the pancreatic cancer comprises at least one selected from the group consisting of pancreatic adenocarcinoma, acinar cell carcinoma, cystadenocarcinoma, pancreatoblastoma, adenosquamous carcinoma, signet ring cell carcinoma, hepatoid carcinoma, colloid carcinoma, undifferentiated carcinoma, undifferentiated carcinoma with osteoclast-like giant cells, solid pseudopapillary tumor, and pancreatic mucinous cystic neoplasm.

In certain embodiments, the at least one immunotherapeutic agent comprises an immune checkpoint inhibitor. In certain embodiments, the immune checkpoint inhibitor is selected from the group consisting of a PD-l inhibitor, PD-L1 inhibitor, CTLA-4 inhibitor, TIGIT inhibitor, LAG3 inhibitor, IDO(l/2) inhibitor, and B7-H3 inhibitor. In certain embodiments, the PD-L1 inhibitor comprises avelumab.

In certain embodiments, the estrogen and/or GPER agonist, and the at least one additional anticancer agent, are co-administered to the subject. In certain embodiments, the estrogen and/or GPER agonist, and the at least one additional anticancer agent, are co- formulated.

In certain embodiments, the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, are co-administered to the subject. In certain embodiments, the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, are co- formulated.

In certain embodiments, administering the estrogen and/or GPER agonist, and the at least one additional anticancer agent, to the subject allows for greater treatment efficacy than administering the estrogen and/or GPER agonist, and the at least one additional anticancer agent, separately.

In certain embodiments, administering the estrogen and/or GPER agonist, and the at least one additional anticancer agent, to the subject enhances the anti-cancer activity of the estrogen and/or GPER agonist, and the at least one additional anticancer agent, in the subject.

In certain embodiments, administering the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, to the subject allows for greater treatment efficacy than administering the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, separately.

In certain embodiments, administering the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, to the subject enhances the anti-cancer activity of the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, in the subject.

In certain embodiments, the estrogen and/or GPER agonist is/are the only anticancer agent administered to the subject.

In certain embodiments, the estrogen and/or GPER agonist are/is the only anticancer agent administered to the subject in an amount sufficient to treat or prevent the pancreatic cancer in the subject.

In certain embodiments, the estrogen and/or GPER agonist are/is the only anticancer agent administered to the subject in an amount sufficient to treat or prevent the MCC in the subject.

In certain embodiments, the estrogen comprises at least one selected from the group consisting of estrone (El), estradiol (E2), estriol (E3), estetrol (E4), l7P-estradiol, 27- hydroxycholesterol, dehydroepiandrosterone (DHEA), 7-oxo-DHEA, 7a-hydroxy-DHEA, l6a-hydroxy-DHEA, 7P-hydroxyepiandrosterone, A⁴-androstenedione, A⁵-androstenediol, 3a-androstanediol, 3P-androstanediol, 2-hydroxy estrone, l6-hydroxyestrone, estradiol cypionate, estradiol valerate, estradiol acetate, estradiol benzoate, ethinyl estradiol (EE), mestranol, moxestrol, quinestrol, diethylstilbestrol benzestrol, dienestrol, dienestrol acetate, diethylstilbestrol dipropionate, fosfestrol, hexestrol, methestrol dipropionate, xenoestrogens, phytoestrogens, and mycoestrogens, or a salt, solvate, tautomer, enantiomer or

diastereoisomer thereof.

In certain embodiments, the GPER agonist comprises at least one selected from the group consisting of G-l, tamoxifen, fulvestrant, and raloxifene, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof.

In certain embodiments, the GPER agonist comprises at least one selected from the group consisting of:

a molecule of formula

wherein in (I):

Ri is selected from the group consisting of =0, =N-OH, =N-NHC(=0)(p-methoxy phenyl), =N-NHC(=0)CH(OMe)phenyl, and =N-NH(5-iodo-pyrid-2-yl);

R₂ is selected from the group consisting of C₁-C₄ alkyl and C₁-C₄ haloalkyl;

bond a is a single or double bond, such that:

if bond a is a double bond, R₃ and R₄ are H, and

if bond a is a single bond, R₃ is selected from the group consisting of H, -OH, -OAc, and halo; R₄ is selected from the group consisting of H, -OH, -OAc, and -S(o- nitrophenyl); or R₃ and R₄ combine to form a diradical selected from the group consisting of -CH₂-, -0CH₂0-, -0CH(CH₃)0-, and -0C(CH₃)₂0-;

R₅ is selected from the group consisting of H, benzyl, C₁-C₄ alkyl, and acetyl;

s is selected from the group consisting of H, halo, -N0₂, C₁-C₄ alkyl, -CºCH, -CºC- Si(CH₃)₃ (or -CºC-TMS), -O-benzyl, -OH, -OAc, C₁-C₄ alkoxy, -COOH, and -COO(Ci-C₄ alkyl);

R₇ is selected from the group consisting of H, halo, -N0₂ C₁-C₄ alkyl, -OH, -OAc, and C₁-C₄ alkoxy;

Rs is selected from the group consisting of H, halo, -N0₂, C₁-C₄ alkyl, -O-benzyl, -N(R)(R), - SR, -COOH, -COO(Ci-C₄ alkyl), -OH, -OAc, C₁-C₄ alkoxy, 3 -thietyl-m ethoxy, - S0₂(morpholino), and -OCH₂CH=CH₂, wherein each occurrence of R is independently selected from the group consisting of H and C₁-C₄ alkyl;

R₉ is selected from the group consisting of H, halo, -N0₂, C₁-C₄ alkyl, -OH, -OAc, and C₁-C₄ alkoxy,

or R₈ and R₉ combine to form a diradical selected from the group consisting of -0CH₂0-, - 0CH(CH₃)0-, -0C(CH₃)₂0-, -0(CH₂)₂0-, -0-CH=CH-, and -CH=CH-0-;

Rio is selected from the group consisting of H, C₁-C₄ alkyl, and halo;

wherein each occurrence of benzyl is independently optionally substituted with at least one group selected from the group consisting of C₁-C₄ alkyl, -OH, C₁-C₄ alkoxy, halo, and -N0₂; and

a molecule of formula

, wherein in (II):

Ri is selected from the group consisting of =0 and =N-OH;

R₂ is C₁-C₄ alkyl;

R₅ is selected from the group consisting of H, benzyl and C₁-C₄ alkyl;

R₈ and R₉ are independently selected from the group consisting of H and C₁-C₄ alkoxy, or R₈ and R₉ combine to form a diradical selected from the group consisting of -0CH₂0-, - 0CH(CH₃)0- and -0C(CH₃)₂0-,

or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, or any mixtures thereof.

a molecule of Formula

wherein in (1-1):

Ri is selected from the group consisting of =0 and =N-OH;

R₂ is C₁-C₄ alkyl;

bond a is a single or double bond, such that: if bond a is a double bond, R₃ and R₄ are H, and

if bond a is a single bond, R₃ and R4 are independently selected from the group consisting of H and -OH, or R₃ and R4 combine to form a diradical selected from the group consisting of -0CH₂0-, -0CH(CH₃)0- and -0C(CH₃)₂0-;

R₅ is selected from the group consisting of H, benzyl and C₁-C₄ alkyl;

s is selected from the group consisting of H and halo;

R₈ and R9 are independently selected from the group consisting of H and C1-C4 alkoxy, or R₈ and R9 combine to form a diradical selected from the group consisting of -0CH₂0-, - 0CH(CH₃)0- and -0C(CH₃)₂0-;

and

a molecule of formula

wherein in (II):

Ri is selected from the group consisting of =0 and =N-OH;

R₂ is C₁-C₄ alkyl;

R₅ is selected from the group consisting of H, benzyl and C₁-C₄ alkyl;

R₇ and R₈ are independently selected from the group consisting of H and C1-C4 alkoxy, or R₇ and R₈ combine to form a diradical selected from the group consisting of -OCH₂0-, - OCH(CH₃)0- and -0C(CH₃)₂0-,

In certain embodiments, the GPER agonist comprises at least one selected from the group consisting of: G-l; CMPD1 (rel-l-((3aS,4R,9bR)-4-(benzo[d][l,3]dioxol-5-yl)- 3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-l-one); CMPD2 (rel-l- ((3aS,4R,9bR)-4-(6-bromobenzo[d][l,3]dioxol-5-yl)-5-methyl-3a,4,5,9b-tetrahydro-3H- cyclopenta[c]quinolin-8-yl)ethan-l-one); CMPD3 (rel-l-((3aS,4R,9bR)-4-(6- bromobenzo[d][l,3]dioxol-5-yl)-2,3,3a,4,5,9b-hexahydro-lH-cyclopenta[c]quinolin-8- yl)ethan-l-one); CMPD4 (rel-l-((3aS,4R,9bR)-5-benzyl-4-(6-bromobenzo[d][l,3]dioxol-5- yl)-3 a, 4, 5,9b-tetrahy dro-3H-cyclopenta[c]quinolin-8-yl)ethan- 1 -one); CMPD5 (rel- 1 - ((3aS,4R,9bR)-4-(2-bromophenyl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8- yl)ethan- 1 -one); CMPD6 (rel- 1 -((3 aS,4R,9bR)-4-(6-bromobenzo[d] [ 1 ,3 ]dioxol-5-yl)- 3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-l-one oxime); CMPD7 (rel-l- ((3aS,4R,9bR)-4-(2-bromo-4,5-dimethoxyphenyl)-3a,4,5,9b-tetrahydro-3H- cyclopenta[c]quinolin-8-yl)ethan-l-one); CMPD8 (rel-l-((3aS,4R,9bR)-4-(6- chlorobenzo[d][l,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-l- one); CMPD9 (rel- 1 -((6R,6aS,7aS, 1 OaR, 10bR)-6-(6-bromobenzo[d] [ 1 ,3]dioxol-5-yl)-9,9- dimethyl-O^a^ aOOa^Ob-hexahydro-SH-fl^jdioxolo^^S’^^jcyclopentafl^-cjquinolin- 2-yl)ethan-l-one); CMPD10 (rel-l-((lR,2S,3aS,4R,9bR)-4-(6-bromobenzo[d][l,3]dioxol-5- yl)-l,2-dihydroxy-2,3,3a,4,5,9b-hexahydro-lH-cyclopenta[c]quinolin-8-yl)ethan-l-one); CMPD11 (rel-l-((3aS,4R,9bR)-4-(2-bromo-4,5-dimethoxyphenyl)-2,3,3a,4,5,9b-hexahydro- lH-cyclopenta[c]quinolin-8-yl)ethan-l-one); and CMPD12 (rel-l-((4S,5aS,6R,l laR)- d^^a^^ lO la-hexahydro-d^-methanofl^jdioxolofd^S’A^jbenzofl^-cjacridin-S- yl)ethan-l-one), or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, or any mixtures thereof.

In certain embodiments, the subject is further administered at least one additional anticancer treatment. In certain embodiments, the at least one additional anticancer treatment comprises chemotherapy, an engineered chimeric antigen receptor (CAR) T-cell, and/or radiation therapy. In certain embodiments, the chemotherapy is selected from the group consisting of a histone deacetylase inhibitor (HD AC), temozolomide, dacarbazine (DTIC), vemurafenib, dabrafenib and trametinib.

In certain embodiments, the estrogen and/or GPER agonist is/are independently administered to the subject by at least one administration route selected from the group consisting of inhalational, oral, rectal, vaginal, parenteral, topical, transdermal, pulmonary, intranasal, buccal, ophthalmic, intrathecal, intracranial, and intravenous.

In certain embodiments, the subject is a mammal. In certain embodiments, the mammal is human.

BRIEF DESCRIPTION OF THE DRAWINGS

The following detailed description of specific embodiments of the invention will be better understood when read in conjunction with the appended drawings. For the purpose of illustrating the invention, certain embodiments are depicted in the drawings. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.

FIGs. 1 A-1K illustrate results that indicate that GPER signaling is tumor suppressive in pancreatic ductal adenocarcinoma (PDAC). FIG. 1 A: Western blot of a dose response treatment of 2838c3 PD AC cells with G-l. FIG. 1B: Western blot of 2838c3 PDAC cells treated with a pulse or constant 500 nM dose of G-l. FIG. 1C: Proliferation assay of several PDAC lines treated with 500 nM G-l, * denotes p < 0.05, error bars = ± s.d. FIGs. 1D-1G: Western blot for p-RB, RB, and c-Myc on several PDAC cell lines. FIG. 1H: Experimental outline of 2838c3 PD AC -bearing mice treated with vehicle or G-l, as well as isotype antibody control (2 A3) or aPD-l antibody. Treatment was started at day 7 after tumors were palpable. n = 5 per group. FIG. II: Tumor volumes after G-l treatment. FIG. 1J: Tumor volumes over time of treatment groups. FIG. 1K: Survival curve of mice treated with vehicle or G-l, as well as aPD-l antibody or isotype antibody control (2A3). Significance between groups by the Log-Rank (Mantel-Cox) test is listed in the table in FIG. 1K.

FIG. 2 comprises a graph illustrating proliferation of Merkel cell carcinoma cells in the presence of varying concentrations of G-l.

DETAILED DESCRIPTION OF THE INVENTION

In one aspect, the present invention relates to the unexpected discovery that estrogen and/or small molecule G-protein coupled receptor (GPCR) agonists (such as selective G protein-coupled estrogen receptor (GPER) agonists) can be used in combination with immunotherapy to treat and/or prevent pancreatic cancer, such as but not limited to K-Ras- driven pancreatic cancer. In certain embodiments, the combination of estrogen/GPCR agonists and immunotherapy is synergistic in nature, allowing for higher efficacy than the sum of thr efficacies that would be obtained with the separate administration of each therapeutic agent. In other embodiments, the combination of estrogen/GPCR agonists and immunotherapy allows for significant cure rates, while the individual administration of each therapeutic agent does not provide significant (or even measurable) increase in survival.

Pancreatic cancer can be subdivided in exocrine pancreatic cancers (which affect areas of the pancreas that produce digestive enzymes) and endocrine pancreatic cancers (which affect areas of the pancreas that produce hormones, and are rather rare as compared to the exocrine subtype). Exocrine pancreatic cancers comprise pancreatic adenocarcinomas (which include invasive or ductal variants), acinar cell carcinomas, cystadenocarcinomas, pancreatoblastomas, adenosquamous carcinomas, signet ring cell carcinomas, hepatoid carcinomas, colloid carcinomas, undifferentiated carcinomas, undifferentiated carcinomas with osteoclast-like giant cells, solid pseudopapillary tumors, and pancreatic mucinous cystic neoplasms.

In certain embodiments, subjects at increased risk of pancreatic cancer, such as but not limited to K-Ras-driven pancreatic cancer, can benefit from treatment with estrogen and/or a GPCR agonist (such as a selective GPER agonist). In other embodiments, a general subject can benefit from treatment with estrogen and/or a GPCR agonist (such as a selective GPER agonist), which acts as a preventive or maintenance treatment against future occurrences of pancreatic cancer, such as but not limited to K-Ras-driven pancreatic cancer. In yet other embodiments, current pancreatic cancer patients, such as but not limited to K- Ras-driven pancreatic cancer patients, can benefit from treatment with estrogen and/or a GPCR agonist (such as a selective GPER agonist), either alone and/or in combination with immunotherapies.

In another aspect, the present invention relates to the unexpected discovery that estrogen and/or small molecule GPCR agonists can be used to treat and/or prevent Merkel cell carcinoma (MCC).

MCC is a rare and highly aggressive skin cancer, which, in most cases, is caused by the Merkel cell polyomavirus (MCV). MCC is also known as cutaneous APUDoma, primary neuroendocrine carcinoma of the skin, primary small cell carcinoma of the skin, or trabecular carcinoma of the skin. MCC occurs most often in Caucasians between 60 and 80 years of age, and its rate of incidence is about twice as high in males as in females. There are roughly 1,500 new cases of MCC diagnosed each year in the ETS. MCC is considered to be a form of neuroendocrine tumor. While patients with a small tumor (< 2 cm) that has not yet metastasized to regional lymph nodes have an expected 5-year survival rate of more than 80%, once a lesion has metastasized regionally, the rate drops to about 50%. . Up to 50% of patients that have been seemingly treated successfully (i.e. that initially appear cancer-free) subsequently suffer a recurrence of their disease.

Immunosuppression can profoundly increase the odds of developing MCC. That cancer occurs 30 times more often in people with chronic lymphocytic leukemia and 13.4 times more often in people with advanced HIV as compared to the general population. Solid organ transplant recipients have a lO-fold increased risk for MCC compared to the general population.

As demonstrated herein, GPER agonists have significant activity against MCC. The in vitro data shows that MCC responds to GPER agonist at the same concentrations that are effective against melanoma, pancreas, and lung cancer cells. MCC was recently shown to be highly responsive to anti -PD- 1 therapy (50% cured and 50% progress). In certain embodiments, estrogen and/ GPCR agonists can be used in combination with immunotherapy to treat and/or prevent MCC. In other embodiments, that combination provides better outcomes (expressed as, for example, life expectancy and/or efficacy) that are superior to those observed with immunotherapy alone.

In certain embodiments, subjects at increased risk of MCC can benefit from treatment with estrogen and/or a GPCR agonist (such as a selective GPER agonist). In other embodiments, a general subject can benefit from treatment with estrogen and/or a GPCR agonist (such as a selective GPER agonist), which acts as a preventive or maintenance treatment against future occurrences of MCC. In yet other embodiments, current MCC patients can benefit from treatment with estrogen and/or a GPCR agonist (such as a selective GPER agonist), either alone and/or in combination with immunotherapies.

Certain compounds, compositions, and methods recited herein are described in PCT International Application No. PCT/US2017/035278, which is incorporated herein in its entirety by reference.

Definitions

As used herein, each of the following terms have the meaning associated with it in this section.

ETnless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics and chemistry are those well-known and commonly employed in the art.

The articles“a” and“an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example,“an element” means one element or more than one element.

As used herein, the term“about” will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. As used herein when referring to a measurable value such as an amount, a temporal duration, and the like, the term “about” is meant to encompass variations of ±20% or ±10%, ±5%, ±1%, or ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

As used herein, the term“alkoxy” employed alone or in combination with other terms means, unless otherwise stated, an alkyl group having the designated number of carbon atoms, as defined elsewhere herein, connected to the rest of the molecule via an oxygen atom, such as, for example, methoxy, ethoxy, l-propoxy, 2-propoxy (or isopropoxy) and the higher homologs and isomers. A specific example is (Ci-C₃)alkoxy, such as, but not limited to, ethoxy and methoxy. As used herein, the term“alkyl” by itself or as part of another substituent means, unless otherwise stated, a straight or branched chain hydrocarbon having the number of carbon atoms designated (i.e., Ci-Cio means one to ten carbon atoms) and includes straight, branched chain, or cyclic substituent groups. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, /f/V-butyl, pentyl, neopentyl, hexyl, and cyclopropylmethyl. A specific embodiment is (Ci-C₄)alkyl, such as, but not limited to, ethyl, methyl, isopropyl, n- butyl, isobutyl, t-butyl, and cyclopropylmethyl.

A“disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate. In contrast, a“disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal’s state of health.

“Effective amount” or“therapeutically effective amount” are used interchangeably herein, and refer to an amount of a compound, formulation, material, or composition, as described herein effective to achieve a particular biological result or provides a therapeutic or prophylactic benefit. Such results may include, but are not limited to, anti-tumor activity as determined by any means suitable in the art.

As used herein“endogenous” refers to any material from or produced inside an organism, cell, tissue or system.

“Estrogen” or“oestrogen” as used herein refers to any substance, natural or synthetic (including analogues and derivatives of estrogen), that mimics the effect of the natural hormone, estrogen. Types of estrogen include, but are not limited to, estrone (El), estradiol (E2), estriol (E3), estetrol (E4), l7P-estradiol, 27-hydroxy cholesterol,

dehydroepiandrosterone (DHEA), 7-oxo-DHEA, 7a-hydroxy-DHEA, l6a-hydroxy-DHEA, 7P-hydroxyepiandrosterone, A⁴-androstenedione, A⁵-androstenediol, 3a-androstanediol, 3b- androstanediol, 2-hydroxyestrone, l6-hydroxyestrone, estradiol cypionate, estradiol valerate, estradiol acetate, estradiol benzoate, ethinyl estradiol (EE), mestranol, moxestrol, quinestrol, diethylstilbestrol benzestrol, dienestrol, dienestrol acetate, diethylstilbestrol dipropionate, fosfestrol, hexestrol, methestrol dipropionate, xenoestrogens, phytoestrogens, and/or mycoestrogens.

As used herein, the term“halo” or“halogen” alone or as part of another substituent refers to, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

As used herein, the term“GPCR” refers to G-protein coupled receptor. As used herein, the term“GPER” refers to a G protein-coupled estrogen receptor, which is a type of GPCR.

As used herein, an“instructional material” includes a publication, a recording, a diagram, or any other medium of expression which can be used to communicate the usefulness of the compositions and methods of the invention. The instructional material of the kit of the invention may, for example, be affixed to a container which contains the nucleic acid, peptide, and/or composition of the invention or be shipped together with a container which contains the nucleic acid, peptide, and/or composition. Alternatively, the instructional material may be shipped separately from the container with the intention that the instructional material and the compound be used cooperatively by the recipient.

As used herein,“immune checkpoint inhibitor” refers to a drug (such as a small molecule, peptide and/or antibody) that triggers an immune system attack on cancer cells. Examples of immune checkpoint inhibitors include, but are not limited to, antibodies, PD-l inhibitors (i.e. Pembrolizumab, Nivolumab, anti-PD-l), PD-L1 inhibitors (i.e. Avelumab, Atezolizumab, anti-PD-Ll), CTLA-4 inhibitors (i.e. Ipilimumab, anti-B7-l/B7-2, anti- CTLA-4), Indoleamine (2, 3)-di oxygenase (IDO 1/2) inhibitors, B7 homolog 3 (B7-H3) inhibitors, lymphocyte activation gene 3 (LAG3) inhibitors, and TIGIT (T cell

immunoreceptor with Ig and ITIM domains) targeting antibodies and agents.

By the term“modified” as used herein, is meant a changed state or structure of a molecule or cell of the invention. Molecules may be modified in many ways, including chemically, structurally, and functionally. Cells may be modified through the introduction of nucleic acids.

By the term“modulating,” as used herein, is meant mediating a detectable increase or decrease in the level of a response in a subject compared with the level of a response in the subject in the absence of a treatment or compound, and/or compared with the level of a response in an otherwise identical but untreated subject. The term encompasses perturbing and/or affecting a native signal or response thereby mediating a beneficial therapeutic response in a subject, preferably, a human.

“Parenteral” administration of an immunogenic composition includes, e.g., subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, or infusion techniques.

As used herein, the term“pharmaceutical composition” or“composition” refers to a mixture of at least one compound useful within the invention with other chemical

components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, and/or excipients. The pharmaceutical composition facilitates administration of the compound to an organism. Multiple techniques of administering a compound exist in the art including, but not limited to: intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary, intracranial, transdermal and topical administration. In certain embodiments, the administration comprises topical administration.

As used herein, the term“pharmaceutically acceptable” refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the composition, and is relatively non-toxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

As used herein, the term“pharmaceutically acceptable carrier” means a

pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the invention within or to the subject such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be“acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the invention, and not injurious to the subject. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as com starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil;

glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid;

pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. As used herein,“pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound useful within the invention, and are physiologically acceptable to the subject. Supplementary active compounds may also be incorporated into the compositions. The“pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound useful within the invention.

Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of the invention are known in the art and described, for example in Remington’s Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.

As used herein, the language“pharmaceutically acceptable salt” refers to a salt of the administered compound prepared from pharmaceutically acceptable non-toxic acids and/or bases, including inorganic acids, inorganic bases, organic acids, inorganic bases, solvates (including hydrates) and clathrates thereof.

As used herein, a“pharmaceutically effective amount,”“therapeutically effective amount” or“effective amount” of a compound is that amount of compound that is sufficient to provide a beneficial effect to the subject to which the compound is administered.

The term“prevent,”“preventing” or“prevention” as used herein means avoiding or delaying the onset of symptoms associated with a disease or condition in a subject that has not developed such symptoms at the time the administering of an agent or compound commences. Disease, condition and disorder are used interchangeably herein.

By the term“specifically bind” or“specifically binds” as used herein is meant that a first molecule preferentially binds to a second molecule (e.g., a particular receptor or enzyme), but does not necessarily bind only to that second molecule.

The term“subject” is intended to include living organisms in which an immune response can be elicited (e.g., mammals). A“subject” or“patient,” as used therein, may be a human or non-human mammal. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and murine mammals. Preferably, the subject is human.

As used herein, a“substantially purified” cell is a cell that is essentially free of other cell types. A substantially purified cell also refers to a cell that has been separated from other cell types with which it is normally associated in its naturally occurring state. In certain embodiments, a population of substantially purified cells refers to a homogenous population of cells. In other embodiments, this term refers simply to cell that have been separated from the cells with which they are naturally associated in their natural state. In yet other embodiments, the cells are cultured in vitro. In yet other embodiments, the cells are not cultured in vitro.

The term“therapeutic” as used herein means a treatment and/or prophylaxis. A therapeutic effect is obtained by suppression, remission, or eradication of a disease state. As used herein,“topical administration” or“topical application” refers to a medication applied to body surfaces such as the skin or mucous membranes.

To“treat” a disease as the term is used herein, means to reduce the frequency or severity of at least one sign or symptom of a disease or disorder experienced by a subject. As used herein, the term“treatment” or“treating” is defined as the application or administration of a therapeutic agent, i.e., a composition useful within the invention (alone or in combination with another pharmaceutical agent), to a subject, or application or administration of a therapeutic agent to an isolated tissue or cell line from a subject (e.g., for diagnosis or ex vivo applications), who has a disease or disorder, a symptom of a disease or disorder or the potential to develop a disease or disorder, with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve or affect the disease or disorder, the symptoms of the disease or disorder or the potential to develop the disease or disorder. Such treatments may be specifically tailored or modified, based on knowledge obtained from the field of pharmacogenomics.

As used herein, the term“UV” refers to ultraviolet.

Throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example,

1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

Compounds and Compositions

In one aspect, the present invention contemplates the use of estrogen and/or other small molecules that bind to and activate a GPCR, such as the G protein-coupled estrogen receptor GPER. In certain embodiments, this induces cell signaling events that increase the differentiation state of the tumor cell. In other embodiments, this slows tumor cell proliferation, slows overall tumor growth, and renders tumor cells more visible to immune cells and/or susceptible to immunotherapy. In yet other embodiments, the present invention contemplates other molecules that serve as agonists of other GPCRs. Such molecules induce cell differentiation by engaging many of the same downstream pathways that are activated by GPER.

In certain embodiments, the compound, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, is at least one compound of formula (I):

wherein in (I):

R₂ is selected from the group consisting of C₁-C₄ alkyl and Ci-C₄ haloalkyl (such as, but not limited to, -CF₃);

bond a is a single or double bond, such that:

if bond a is a double bond, R₃ and R* are H, and

if bond a is a single bond, R₃ is selected from the group consisting of H, -OH, -OAc, and halo; * is selected from the group consisting of H, -OH, -OAc, and -S(o-nitrophenyl); or R₃ and R₄ combine to form a diradical selected from the group consisting of -CH₂-, -0CH₂0-, -0CH(CH₃)0-, and -0C(CH₃)₂0-;

R₅ is selected from the group consisting of H, benzyl, C₄-C₄ alkyl, and acetyl;

s is selected from the group consisting of H, halo, -N0₂, C₄-C₄ alkyl, -CºCH, -CºC- Si(CH₃)₃ (or -CºC-TMS), -O-benzyl, -OH, -OAc, C₄-C₄ alkoxy, -COOH, and - COO(C C₄ alkyl);

R₇ is selected from the group consisting of H, halo, -N0_2. C₄-C₄ alkyl, -OH, -OAc, and C₄-C₄ alkoxy;

Rs is selected from the group consisting of H, halo, -N0₂, C₄-C₄ alkyl, -O-benzyl, -N(R)(R), - SR, -COOH, -COO(Ci-C alkyl), -OH, -OAc, Ci-C alkoxy, 3 -thietyl-m ethoxy, - S0₂(morpholino), and -OCH₂CH=CH₂, wherein each occurrence of R is

independently selected from the group consisting of H and C₄-C₄ alkyl;

R₉ is selected from the group consisting of H, halo, -N0₂, C₄-C₄ alkyl, -OH, -OAc, and C₄-C₄ alkoxy,

or R₈ and R₉ combine to form a diradical selected from the group consisting of - 0CH₂0-, -0CH(CH₃)0-, -0C(CH₃)₂0-, -0(CH₂)₂0-, -0-CH=CH-, and -CH=CH-0-;

Rio is selected from the group consisting of H, C₄-C₄ alkyl, and halo; wherein each occurrence of benzyl is independently optionally substituted with at least one group selected from the group consisting of C₁-C₄ alkyl, -OH, Ci-C₄ alkoxy, halo, and -N0_2.

In certain embodiments, the compound, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, is at least one compound of formula (1-1):

wherein in (1-1):

Ri is selected from the group consisting of =0 and =N-OH;

R₂ is Ci-C₄ alkyl;

bond a is a single or double bond, such that:

if bond a is a double bond, R₃ and R* are H, and

if bond a is a single bond, R₃ and * are independently selected from the group consisting of H and -OH, or R₃ and R₄ combine to form a diradical selected from the group consisting of -OCH₂0-, -OCH(CH₃)0- and - 0C(CH₃)₂0-;

R₅ is selected from the group consisting of H, benzyl and C₄-C₄ alkyl;

s is selected from the group consisting of H and halo;

R₈ and R₉ are independently selected from the group consisting of H and C₄-C₄ alkoxy, or R₈ and R₉ combine to form a diradical selected from the group consisting of -OCH₂0-, - OCH(CH₃)0- and -0C(CH₃)₂0-.

In certain embodiments, the compound is not G-l. In other embodiments, the compound is G-l.

In certain embodiments, Ri is =0. In other embodiments, Ri is =N-OH.

In certain embodiments, R₂ is selected from the group consisting of methyl, trifluoromethyl, ethyl, l-propyl, and 2-propyl. In other embodiments, R₂ is methyl or trifluoromethyl.

In certain embodiments, bond a is a double bond, and R₃ and R₄ are H.

In certain embodiments, bond a is a single bond, and R₃ and R are independently selected from the group consisting of H and -OH. In other embodiments, bond a is a single bond, and R₃ and R₄ are H. In yet other embodiments, bond a is a single bond, and R₃ and R₄ are -OH. In yet other embodiments, bond a is a single bond, and R₃ and R₄ are -OH and cis to each other. In yet other embodiments, bond a is a single bond, and R₃ and R₄ are -OH and trans to each other.

In certain embodiments, bond a is a single bond, and R₃ and R₄ combine to form - 0C(CH₃)₂0-.

In certain embodiments, R₅ is selected from the group consisting of H, benzyl, methyl, ethyl, 1 -propyl and 2-propyl. In certain embodiments, R₅ is selected from the group consisting of H, benzyl and methyl.

In certain embodiments, ₅ is selected from the group consisting of H, F, Cl, Br and I. In certain embodiments, R₅ is selected from the group consisting of H, Cl and Br.

In certain embodiments, R₈ and R₉ are independently selected from the group consisting of H and C₁-C₄ alkoxy. In other embodiments, R₈ and R₉ are independently selected from the group consisting of H, methoxy, ethoxy, l-propoxy and 2-propoxy. In yet other embodiments, R₈ and R₉ are independently selected from the group consisting of H and methoxy.

In certain embodiments, R₈ and R₉ combine to form a diradical selected from the group consisting of -0CH₂0-, -0(CH₂)₂0-, -0CH(CH₃)0- and -0C(CH₃)₂0-.

In certain embodiments, the compound, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, is at least one selected from the group consisting of:

In certain embodiments, the compound is Al. In other embodiments, the compound is A2. In yet other embodiments, the compound is A3. In yet other embodiments, the compound is A4. In yet other embodiments, the compound is A5. In yet other embodiments, the compound is A6. In yet other embodiments, the compound is A7. In yet other embodiments, the compound is A8. In yet other embodiments, the compound is A9. In yet other embodiments, the compound is A10. In yet other embodiments, the compound is Al 1. In yet other embodiments, the compound is A12. In yet other embodiments, the compound is A13. In yet other embodiments, the compound is A14. In yet other embodiments, the compound is A15.

In certain embodiments, the compound is not Al. In other embodiments, the compound is not A2. In yet other embodiments, the compound is not A3. In yet other embodiments, the compound is not A4. In yet other embodiments, the compound is not A5. In yet other embodiments, the compound is not A6. In yet other embodiments, the compound is not A7. In yet other embodiments, the compound is not A8. In yet other embodiments, the compound is not A9. In yet other embodiments, the compound is not A10. In yet other embodiments, the compound is not All. In yet other embodiments, the compound is not A12. In yet other embodiments, the compound is not A13. In yet other embodiments, the compound is not A14. In yet other embodiments, the compound is not A15.

In certain embodiments, the compound is A16. In other embodiments, the compound is A17. In yet other embodiments, the compound is A18. In yet other embodiments, the compound is A19. In yet other embodiments, the compound is A20. In yet other

embodiments, the compound is A21. In yet other embodiments, the compound is A22. In yet other embodiments, the compound is A23. In yet other embodiments, the compound is A24. In yet other embodiments, the compound is A25. In yet other embodiments, the compound is A26. In yet other embodiments, the compound is A27. In yet other embodiments, the compound is A28. In yet other embodiments, the compound is A29. In yet other

embodiments, the compound is A30. In yet other embodiments, the compound is A31. In yet other embodiments, the compound is A32. In yet other embodiments, the compound is A33. In yet other embodiments, the compound is A34. In yet other embodiments, the compound is A35. In yet other embodiments, the compound is A36. In yet other embodiments, the compound is A37. In yet other embodiments, the compound is A38. In yet other embodiments, the compound is A39. In yet other embodiments, the compound is A40. In yet other embodiments, the compound is A41. In yet other embodiments, the compound is A42. In yet other embodiments, the compound is A43. In yet other embodiments, the compound is A44. In yet other embodiments, the compound is A45. In yet other embodiments, the compound is A46. In yet other embodiments, the compound is A47. In yet other

embodiments, the compound is A48. In yet other embodiments, the compound is A49. In yet other embodiments, the compound is A50. In yet other embodiments, the compound is A51. In yet other embodiments, the compound is A52. In yet other embodiments, the compound is A53. In yet other embodiments, the compound is A54. In yet other embodiments, the compound is A55. In yet other embodiments, the compound is A56. In yet other

embodiments, the compound is A57. In yet other embodiments, the compound is A58. In yet other embodiments, the compound is A59. In yet other embodiments, the compound is A60. In yet other embodiments, the compound is A61. In yet other embodiments, the compound is A62. In yet other embodiments, the compound is A63. In yet other embodiments, the compound is A64. In yet other embodiments, the compound is A65. In yet other

embodiments, the compound is A66. In yet other embodiments, the compound is A67. In yet other embodiments, the compound is A68. In yet other embodiments, the compound is A69. In yet other embodiments, the compound is A70. In yet other embodiments, the compound is A71. In yet other embodiments, the compound is A72. In yet other embodiments, the compound is A73. In yet other embodiments, the compound is A74. In yet other

embodiments, the compound is A75. In yet other embodiments, the compound is A76. In yet other embodiments, the compound is A77. In yet other embodiments, the compound is A78. In yet other embodiments, the compound is A79. In yet other embodiments, the compound is A80. In yet other embodiments, the compound is A81. In yet other embodiments, the compound is A82. In yet other embodiments, the compound is A83. In yet other

embodiments, the compound is A84. In yet other embodiments, the compound is A85. In yet other embodiments, the compound is A86. In yet other embodiments, the compound is A87. In yet other embodiments, the compound is A88. In yet other embodiments, the compound is A89. In yet other embodiments, the compound is A90. In yet other embodiments, the compound is A91. In yet other embodiments, the compound is A92. In yet other

embodiments, the compound is A93. In yet other embodiments, the compound is A94. In yet other embodiments, the compound is A95. In yet other embodiments, the compound is A96. In yet other embodiments, the compound is A97. In yet other embodiments, the compound is A98. In yet other embodiments, the compound is A99. In yet other embodiments, the compound is A100. In yet other embodiments, the compound is A101. In yet other embodiments, the compound is A102. In yet other embodiments, the compound is A103. In yet other embodiments, the compound is A104.

In certain embodiments, the compound is not A16. In other embodiments, the compound is not A17. In yet other embodiments, the compound is not A18. In yet other embodiments, the compound is not A19. In yet other embodiments, the compound is not A20. In yet other embodiments, the compound is not A21. In yet other embodiments, the compound is not A22. In yet other embodiments, the compound is not A23. In yet other embodiments, the compound is not A24. In yet other embodiments, the compound is not A25. In yet other embodiments, the compound is not A26. In yet other embodiments, the compound is not A27. In yet other embodiments, the compound is not A28. In yet other embodiments, the compound is not A29. In yet other embodiments, the compound is not A30. In yet other embodiments, the compound is not A31. In yet other embodiments, the compound is not A32. In yet other embodiments, the compound is not A33. In yet other embodiments, the compound is not A34. In yet other embodiments, the compound is not A35. In yet other embodiments, the compound is not A36. In yet other embodiments, the compound is not A37. In yet other embodiments, the compound is not A38. In yet other embodiments, the compound is not A39. In yet other embodiments, the compound is not A40. In yet other embodiments, the compound is not A41. In yet other embodiments, the compound is not A42. In yet other embodiments, the compound is not A43. In yet other embodiments, the compound is not A44. In yet other embodiments, the compound is not A45. In yet other embodiments, the compound is not A46. In yet other embodiments, the compound is not A47. In yet other embodiments, the compound is not A48. In yet other embodiments, the compound is not A49. In yet other embodiments, the compound is not A50. In yet other embodiments, the compound is not A51. In yet other embodiments, the compound is not A52. In yet other embodiments, the compound is not A53. In yet other embodiments, the compound is not A54. In yet other embodiments, the compound is not A55. In yet other embodiments, the compound is not A56. In yet other embodiments, the compound is not A57. In yet other embodiments, the compound is not A58. In yet other embodiments, the compound is not A59. In yet other embodiments, the compound is not A60. In yet other embodiments, the compound is not A61. In yet other embodiments, the compound is not A62. In yet other embodiments, the compound is not A63. In yet other embodiments, the compound is not A64. In yet other embodiments, the compound is not A65. In yet other embodiments, the compound is not A66. In yet other embodiments, the compound is not A67. In yet other embodiments, the compound is not A68. In yet other embodiments, the compound is not A69. In yet other embodiments, the compound is not A70. In yet other embodiments, the compound is not A71. In yet other embodiments, the compound is not A72. In yet other embodiments, the compound is not A73. In yet other embodiments, the compound is not A74. In yet other embodiments, the compound is not A75. In yet other embodiments, the compound is not A76. In yet other embodiments, the compound is not A77. In yet other embodiments, the compound is not A78. In yet other embodiments, the compound is not A79. In yet other embodiments, the compound is not A80. In yet other embodiments, the compound is not A81. In yet other embodiments, the compound is not A82. In yet other embodiments, the compound is not A83. In yet other embodiments, the compound is not A84. In yet other embodiments, the compound is not A85. In yet other embodiments, the compound is not A86. In yet other embodiments, the compound is not A87. In yet other embodiments, the compound is not A88. In yet other embodiments, the compound is not A89. In yet other embodiments, the compound is not A90. In yet other embodiments, the compound is not A91. In yet other embodiments, the compound is not A92. In yet other embodiments, the compound is not A93. In yet other embodiments, the compound is not A94. In yet other embodiments, the compound is not A95. In yet other embodiments, the compound is not A96. In yet other embodiments, the compound is not A97. In yet other embodiments, the compound is not A98. In yet other embodiments, the compound is not A99. In yet other embodiments, the compound is not A100. In yet other embodiments, the compound is not A101. In yet other embodiments, the compound is not A102. In yet other embodiments, the compound is not A103. In yet other embodiments, the compound is not A104.

In certain embodiments, the compound of the invention, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, is at least one compound selected from the group

consisting of:

other embodiments, the compound is not A105. In other embodiments, the compound is not A106.

In certain embodiments, the compound of the invention, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof,

other embodiments, the compound is not A107.

In certain embodiments, the compound is at least one selected from the group consisting of:

Gl or G-l (rel-l-[4-(6-bromo-l,3-benzodioxol-5-yl)-3aR,4S,5,9bS-tetrahydro-3H-

cyclopenta[c]quinolin-8-yl]-ethanone)

CMPD4 (rel-l-((3aS,4R,9bR)-5-benzyl-4-(6-bromobenzo[d][l,3]dioxol-5-yl)-3a,4,5,9b-

tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-l-one):

CMPD5 (rel-l-((3aS,4R,9bR)-4-(2-bromophenyl)-3a,4,5,9b-tetrahydro-3H-

cyclopenta[c]quinolin-8-yl)ethan-l-one)

CMPD6 (rel-l-((3aS,4R,9bR)-4-(6-bromobenzo[d][l,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-

cyclopenta[c]quinolin-8-yl)ethan-l-one oxime):

CMPD7 (rel-l-((3aS,4R,9bR)-4-(2-bromo-4,5-dimethoxyphenyl)-3a,4,5,9b-tetrahydro-3H- cyclopenta[c]quinolin-8-yl)ethan-l-one)

CMPD8 (rel-l-((3aS,4R,9bR)-4-(6-chlorobenzo[d][l,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-

cyclopenta[c]quinolin-8-yl)ethan-l-one):

CMPD9 (rel-l-((6R,6aS,7aS,l0aR,l0bR)-6-(6-bromobenzo[d][l,3]dioxol-5-yl)-9,9- dimethyl-6, 6a, 7, 7a, l0a,l0b-hexahydro-5H-[l,3]dioxolo[4’, 5’ :3,4]cyclopenta[l,2-c]quinolin-

2-yl)ethan-l-one)

CMPD10 (rel-l-((lR,2S,3aS,4R,9bR)-4-(6-bromobenzo[d][l,3]dioxol-5-yl)-l,2-dihydroxy- 2,3,3a,4,5,9b-hexahydro-lH-cyclopenta[c]quinolin-8-yl)ethan-l-one):

CMPD11 (rel-l-((3aS,4R,9bR)-4-(2-bromo-4,5-dimethoxyphenyl)-2,3,3a,4,5,9b-hexahydro-

lH-cyclopenta[c]quinolin-8-yl)ethan-l-one)

In certain embodiments, the compound of the invention, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof, is at least one compound of formula (II):

wherein:

Ri is selected from the group consisting of =0 and =N-OH;

R₂ is C₁-C₄ alkyl;

R5 is selected from the group consisting of H, benzyl and C₁-C₄ alkyl;

R₈ and R₉ are independently selected from the group consisting of H and C1-C4 alkoxy, or R₈ and R9 combine to form a diradical selected from the group consisting of -0CH₂0-, - 0CH(CH₃)0- and -0C(CH₃)₂0-.

In certain embodiments, the compound of formula (II) is

CMPD12 (rel-l-((4S,5aS,6R,llaR)-4,5,5a,6,ll,lla-hexahydro-4,6-methano[l,3]

dioxolo^SkS^Jbenzofl^-cJacridin-S-y^ethan-l-one)

In certain embodiments, the compound useful within the methods of the invention, or a salt, solvate, enantiomer or diastereoisomer thereof, is at least one GPER agonist recited in U.S. Patent Application Publications No. US 2008/0167334 and US 2011/0092533, all of which are incorporated herein in their entireties by reference:

wherein:

X is =N-, O, S, or N-R, with the proviso that when X is N-R and R is a bond, N together with R¹ forms a 5- to 7-membered optionally substituted heterocyclic group;

R is a bond, H, -OH, -N0₂, optionally substituted Ci-C₆ hydrocarbyl (such as optionally substituted alkyl), optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted -C(0)-(Ci-C₆) alkyl (amide), optionally substituted -C(0)-0-(Ci-C₆) alkyl (urethane), optionally substituted -C(0)-NH(Ci- C₆) alkyl (urea), optionally substituted -C(0)-N(Ci-C₆)dialkyl, optionally substituted -C(O)- NH(aryl), optionally substituted -C(0)-N(diaryl), optionally substituted -C(O)- NH(heteroaryl), optionally substituted -C(0)-N(diheteroaryl), optionally substituted -C(O)- NH(heterocycle) or optionally substituted -C(0)-N(diheterocycle);

R¹, R² and R⁵ are each independently selected from H, -OH, -N0₂, halogen, Ci-C₆ optionally substituted carboxylic acid group, optionally substituted 0-(Ci-C₆)alkyl, optionally substituted Ci-C₆ hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted -C(0)-(Ci-C₆) alkyl (ketone), optionally substituted -C(0)-0-(Ci-C₆) alkyl (ester), optionally substituted 0-C(0)-(Ci-C₆) alkyl (ester), optionally substituted -C(0)-NH(Ci-C₆) alkyl (urea), optionally substituted - C(0)-N(Ci-C₆)dialkyl, optionally substituted -C(0)-NH(aryl), optionally substituted -C(O)- N(diaryl), optionally substituted -C(0)-NH(heteroaryl), optionally substituted -C(O)- N(diheteroaryl), optionally substituted -C(0)-NH(heterocycle) or optionally substituted - C(0)-N(diheterocycle);

R³ and R⁴ are each independently selected from H, -OH, -N0₂, halogen, Ci-C₆ optionally substituted carboxylic acid group, optionally substituted 0-(Ci-C₆)alkyl, optionally substituted Ci-C₆ hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted -C(0)-(Ci-C₆) alkyl (ketone), optionally substituted -C(0)-0-(Ci-C₆) alkyl (ester), optionally substituted 0-C(0)-(Ci-C₆) alkyl (ester), optionally substituted -C(0)-NH(Ci-C₆) alkyl (urea), optionally substituted - C(0)-N(Ci-C₆)dialkyl, optionally substituted -C(0)-NH(aryl), optionally substituted -C(O)- N(diaryl), optionally substituted -C(0)-NH(heteroaryl), optionally substituted -C(O)- N(diheteroaryl), optionally substituted -C(0)-NH(heterocycle) or optionally substituted - C(0)-N(diheteroaryl) or R³ and R⁴ together form a 5- or 6-membered optionally substituted carbocyclic (which may be saturated or unsaturated), optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclic group;

R⁶ and R⁷ are each independently absent or are selected from H, -OH, -N0₂, halogen, Ci-C₆ optionally substituted carboxylic acid group, optionally substituted 0-(Ci-C₆)alkyl, optionally substituted Ci-C₆ hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted -C(0)-(Ci-C₆) alkyl (ketone), optionally substituted -C(0)-0-(Ci-C₆) alkyl (ester), optionally substituted O-C(O)- (Ci-C₆) alkyl (ester), optionally substituted -C(0)-NH(Ci-C₆) alkyl (urea), optionally substituted -C(0)-N(Ci-C₆)dialkyl, optionally substituted -C(0)-NH(aryl), optionally substituted -C(0)-N(diaryl), optionally substituted -C(0)-NH(heteroaryl), optionally substituted -C(0)-N(diheteroaryl), optionally substituted -C(0)-NH(heterocycle) or optionally substituted -C(0)-N(diheteroaryl), or together R⁶ and R⁷ form a 4-, 5-, 6- or 7- membered optionally substituted carbocyclic (which may be saturated or unsaturated), optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclic group, or a 5- to 9-membered optionally substituted carbocyclic or heterocyclic bicyclic group, with the proviso that R⁷ is not absent when both R⁷ and R⁷ are also absent;

R^6' is absent, H, Ci-C₆ optionally substituted hydrocarbyl group (such as H, C¾ or CH₂CH₃) or together with R⁶ forms =0 ;

R^7' is absent, H, optionally substituted hydrocarbyl group (such as H, C¾ or CH₂CH₃), or together with R⁷ forms =0;

R^7" is absent, H, -OH, halogen, optionally substituted 0-(Ci-C₆)alkyl, optionally substituted Ci-C₆ hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted -C(0)-(Ci-C₆) alkyl (ketone), optionally substituted -C(0)-0-(Ci-C₆) alkyl (ester), optionally substituted 0-C(0)-(Ci-C₆) alkyl (ester), optionally substituted -C(0)-NH(Ci-C₆) alkyl (urea), optionally substituted - C(0)-N(Ci-C₆)dialkyl, optionally substituted -C(0)-NH(aryl), optionally substituted -C(O)- N(diaryl), optionally substituted -C(0)-NH(heteroaryl), optionally substituted -C(O)- N(diheteroaryl), optionally substituted -C(0)-NH(heterocycle) or optionally substituted - C(0)-N(diheteroaryl);

R⁸ is absent (when the carbon to which R⁸ is attached and the carbon to which R⁶ is attached form an optional double bond), H, C¾ or CH₂CH₃;

R¹⁰, R¹¹, R¹² and R¹³ are each independently H, -OH, -NO₂, halogen, Ci-C₆ optionally substituted carboxylic acid group, optionally substituted 0-(Ci-C₆)alkyl, optionally substituted Ci-C₆ hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted -C(0)-(Ci-C₆) alkyl (ketone), optionally substituted -C(0)-0-(Ci-C₆) alkyl (ester), optionally substituted 0-C(0)-(Ci-C₆) alkyl (ester), optionally substituted -C(0)-NH(Ci-C₆) alkyl (urea), optionally substituted - C(0)-N(Ci-C₆)dialkyl, optionally substituted -C(0)-NH(aryl), optionally substituted -C(O)- N(diaryl), optionally substituted -C(0)-NH(heteroaryl), optionally substituted -C(O)- N(diheteroaryl), optionally substituted -C(0)-NH(heterocycle) or optionally substituted - C(0)-N(diheteroaryl);

R¹⁴ is H, -OH, -NO₂, halogen, Ci-C₆ optionally substituted carboxylic acid group, optionally substituted 0-(Ci-C₆)alkyl, optionally substituted Ci-C₆ hydrocarbyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted -C(0)-(Ci-C₆) alkyl (ketone), optionally substituted -C(0)-0-(Ci-C₆) alkyl (ester), optionally substituted 0-C(0)-(Ci-C₆) alkyl (ester), optionally substituted - C(0)-NH(C_I-C₆) alkyl (urea), optionally substituted -C(0)-N(Ci-C₆)dialkyl, optionally substituted -C(0)-NH(aryl), optionally substituted -C(0)-N(diaryl), optionally substituted - C(0)-NH(heteroaryl), optionally substituted -C(0)-N(diheteroaryl), optionally substituted - C(0)-NH(heterocycle) or optionally substituted -C(0)-N(diheterocycle) or together with the carbon to which R⁷ is attached forms a 5-, 6- or 7-membered optionally substituted carbocyclic (which may be saturated or unsaturated), optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclic ring;

Y is optionally substituted (CH₂)_n group where n is 0, 1 or 2, optionally substituted =CH- group, a C=0 group, O, S, optionally substituted N-(Ci-C₆)alkyl, optionally substituted N-aryl, optionally substituted N-heteroaryl, optionally substituted N-heterocycle, optionally substituted N-C(0)-(Ci-C₆)alkyl, optionally substituted N-C(0)-aryl, optionally substituted N-C(0)-heteroaryl, optionally substituted N-C(0)-heterocycle;

Z is optionally substituted (CH₂)_n group where n is 1 or 2, optionally substituted =CH- group, a C=0 group, O, S, optionally substituted N-(Ci-C₆)alkyl, optionally substituted N- aryl, optionally substituted N-heteroaryl, optionally substituted N-heterocycle, optionally substituted N-C(0)-(Ci-C₆)alkyl, optionally substituted N-C(0)-aryl, optionally substituted N-C(0)-heteroaryl, optionally substituted N-C(0)-heterocycle.

In certain embodiments, the compound of the invention, or a salt, solvate, enantiomer or diastereoisomer thereof, is at least one GPER agonist recited in PCT Patent Application No. WO 2016/014847, which is incorporated herein in its entirety by reference:

(iv), wherein in (i)-(iv):

Ring A is an aromatic or heteroaromatic five or six membered ring containing one or more heteroatom such as N, O, or S;

Ri is independently selected from S0₂NH₂, S0₂NR_aR_b, COOH, CONH₂ and

CONR_aR_b, and H. In Ri, each occurrence of R_a and R_b is independently selected from H, alkyl (Ci-C₆), alkenyl (C₂-C₆), alkynyl (C₂-C₆), alkoxy (C₂-C₆), cycloalkyl (C₃-C₇), alkylthio, alkylaryl, and aromatic and hetroaromatic rings. The aromatic and heteroaromatic rings can be further substituted with electron withdrawing and donating groups. R_a and R_b can form a cyclic ring (C₃-C₇) or an aromatic ring optionally containing one or more heteroatoms. Such aromatic rings can be further substituted with electron withdrawing groups such as halogens, -COOH, -CN, -NO₂ and the like, or electron donating groups such as alkyl groups;

R₂ is H, halogen, or a heteroatom such as N, O, or S;

Ring A is an aromatic or heteroaromatic ring (5 or 6 membered);

Ring B is a six membered saturated or aromatic ring containing N at the indicated position. The nitrogen of ring B optionally can be substituted with an alkyl, aryl, or alkaryl substituent;

Ring C is an independently a substituted or unsubstituted carbocyclic ring, bicyclic ring, aromatic ring, fused aromatic rings, or a heteroaromatic ring. Additionally, when it is a carbocyclic ring, it may contain one or more double bonds and one or more heteroatoms such as N, O, or S. It may also have an a-b unsaturated ketone function;

R₃ is selected independently from H, halogen, -OH, CN, (Ci-C₆) alkyl, (C₂-C₆) alkenyl, (C₂-C₆) alkynyl, (Ci-C₆) alkoxy, (C₃-C₇) cycloalkyl, (Ci-C₆) alkylthio, NR_aR_b, R_aR_b or haloalkyl (e.g., CF₃). In R₃, each occurrence of R_a and R_b is independently hydrogen or (Ci-C₆) alkyl, or R_a and R_b form a saturated or unsaturated heterocyclic ring containing from 3-7 ring atoms, which ring may optionally contain another heteroatom selected from N, O, and S, and may be optionally substituted by from 1-3 groups which may be the same or different and are selected from (C₁-C₄) alkyl, phenyl, and benzyl; and m is 1-4;

Rings B and C are in certain embodiments cis fused;

Rings D and B is directly connected or connected through a spacer (C1-C2). When connected directly, they can be cis or trans with respect to the fusion of Ring B and C. Ring D is an aromatic or heteroaromatic ring containing one or more heteroatoms such as N, O, or S. It can be optionally substituted with R₅ and R₆ groups selected independently, or with - ReR₅ or R5R6;

R5 is independently selected from H, halogen, electron donating groups, and electron withdrawing groups such as alkyl, haloalkyl, alkoxy, -N0₂, -SF₅, -CN, and the like;

Re can be NHC(0)0R_c, 0C(0)NHR_c, C(0)0(CH₂)_nR_c, 0C(0)(CH₂)_nR_c, C(0)NHR_c, or NHC(0)R_c, where n = 0-4; or 5 can be alkyl, branched alkyl (C1-C10), alkynyl (C1-C10), a carbocyclic ring, alkenyl (C1-C10), halogen, CN, COOH, CONH₂, -OH, or NH_2. R_c is alkyl, branched alkyl (C1-C10), alkoxy, alkylamino, acyl, alkynyl (Ci-C₈) or alkenyl (Ci-C₆). R₆ can also be X(CH₂)_n-E, wherein X is NH, O, S, CºC, or HC=CH, and n = 0-2, and E is independently a substituted or unsubstituted carbocyclic ring, bicyclic ring, aromatic ring, fused aromatic rings or heteroaromatic ring;

in certain embodiments, Ri is selected from the group consisting of carboxyl, carboxamide, carboxyalkyl, carboxyaryl, cyano, nitro, hydroxyl, sulfonyl, sulfonamide, alkylsulfonamide, arylsulfonamide, alkylsulfonyl, aralkylsulfonamide,

trifluoromethylsulfonamide, trifluorom ethyl sulfonyl carboxamide, and sulfonylcarbamide. In certain embodiments, Ri is sulfonamide, alkylsulfonamide, or arylsulfonamide. R₂ can be a 3-, 4-, 5-, or 6-membered saturated or aromatic carbon ring or ring system optionally containing one or two heteroatoms selected from N, O, and S, the ring or ring system optionally substituted with one or more substituents selected from the group consisting of cyano, halo, acyl, acyloxy, alkyl, alkoxy, heteroalkyl, alkylester, alkylamido, alkylamino, aryl, aryloxy, arylalkyl, arylester, azido, alkylhalo, alkenyl, alkynyl, alkyl ether, nitro, thiohalo, and thiocyano. R₃ can be H, or a Ci-C₅ alkyl or cycloalkyl group optionally substituted with one or more of cyano, nitro, and one or more aromatic or heteroaromatic groups containing N, O, or S. A and D can be independently selected from CH, CH₂, N, and O, and the bond joining them is a single or double bond as appropriate for the selected atoms. X, Y, and Z are independently selected from no atom (i.e., they are absent), CH, C-halogen,

N, O, and S;

in certain embodiments, R₂ is a substituent represented by -R_dR_cRf or by -R_dCOR_eRf; wherein R_d, R_e, and R_f are independently selected from a 3-, 4-, 5-, or 6-membered saturated or aromatic carbon ring or ring system optionally containing one or two heteroatoms selected from N, O, and S, the ring or ring system optionally substituted with one or more substituents selected from the group consisting of cyano, halo, acyl, acyloxy, alkyl, alkoxy, heteroalkyl, alkylester, alkylamido, alkylamino, aryl, aryloxy, arylalkyl, arylester, azido, alkylhalo, alkenyl, alkynyl, alkyl ether, nitro, thiohalo, and thiocyano;

Compounds of the invention may be prepared according to methods described herein, methods known in the art, and/or methods described in certain references, such as but not limited to: PCT Application Publications No. WO 2004/072046 and No. WO 2016/014847; U.S. Application No. 10/511,083; U.S. Patent Application Publications No. US

2008/0167334 and US 2011/0092533; and Burai, et al. , 2010, Org. & Biomol. Chem. 8:2252- 2259; all of which are included herein in their entireties by reference.

The compounds of the invention may possess one or more stereocenters, and each stereocenter may exist independently in either the ( R ) or (S) configuration. In certain embodiments, compounds described herein are present in optically active or racemic forms. The compounds described herein encompass racemic, optically active, regioisomeric and stereoisomeric forms, or combinations thereof that possess the therapeutically useful properties described herein. Preparation of optically active forms is achieved in any suitable manner, including by way of non-limiting example, by resolution of the racemic form with recrystallization techniques, synthesis from optically active starting materials, chiral synthesis, or chromatographic separation using a chiral stationary phase. A compound illustrated herein by the racemic formula further represents either of the two enantiomers or mixtures thereof, or in the case where two or more chiral center are present, all diastereomers or mixtures thereof.

In certain embodiments, the compounds of the invention exist as tautomers. All tautomers are included within the scope of the compounds recited herein.

Compounds described herein also include isotopically labeled compounds wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds described herein include and are not limited to ²H, ¾ ^UC, ¹³C, ¹⁴C, ³⁶Cl, ¹⁸F, ¹²³1, ¹²⁵I, ¹³N, ¹⁵N, ¹⁵0, ¹⁷0, ¹⁸0, ³²P, and ³⁵S. In certain embodiments, substitution with heavier isotopes such as deuterium affords greater chemical stability. Isotopically labeled compounds are prepared by any suitable method or by processes using an appropriate isotopically labeled reagent in place of the non-labeled reagent otherwise employed.

In certain embodiments, the compounds described herein are labeled by other means, including, but not limited to, the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.

In all of the embodiments provided herein, examples of suitable optional substituents are not intended to limit the scope of the claimed invention. The compounds of the invention may contain any of the substituents, or combinations of substituents, provided herein.

In certain embodiments, the invention further provides pharmaceutical compositions comprising at least one compound of the invention and at least one pharmaceutically acceptable carrier. In other embodiments, the pharmaceutical composition is formulated for inhalational, oral, rectal, vaginal, parenteral, topical, transdermal, pulmonary, intranasal, buccal, ophthalmic, intrathecal, intracranial, or intravenous administration.

In certain embodiments, the pharmaceutical compositions of the invention comprise at least one additional anticancer agent and at least one compound of the invention. Examples of additional anticancer agents include, but are not limited to, chemotherapy, and immune checkpoint inihibitors. Non-limiting examples of chemotherapy include, but are not limited to, a HD AC, temozolomide, dacarbazine (DTIC), vemurafenib, dabrafenib and trametinib. Non-limiting examples of checkpoint inhibitors include, but are not limited to, PD-l inhibitors (i.e. Pembrolizumab, Nivolumab, anti-PD-l), PD-L1 inhibitors (i.e., Avelumab, Atezolizumab, anti-PD-Ll), CTLA-4 inhibitors (i.e. Ipilimumab, anti-B7-l/B7-2, anti- CTLA-4), Indoleamine (2, 3)-di oxygenase (IDO 1/2) inhibitors, B7 homolog 3 (B7-H3) inhibitors, lymphocyte activation gene 3 (LAG3) inhibitors, and TIGIT (T cell

immunoreceptor with Ig and ITIM domains) targeting antibodies and agents.

The present invention also pertains to kits useful within any of the methods of the invention described herein. Such kits comprise components useful in any of the methods described herein, one or more containers (e.g., test tube, cell culture dish, cell culture plate, cell culture flask, cell culture bag) for containing a component of any of the embodiments of the invention described elsewhere herein, and instructional materials.

Salts

The compounds described herein may form salts with acids or bases, and such salts are included in the present invention. The term“salts” embraces addition salts of free acids or bases that are useful within the methods of the invention. The term“pharmaceutically acceptable salt” refers to salts that possess toxicity profiles within a range that affords utility in pharmaceutical applications. In certain embodiments, the salts are pharmaceutically acceptable salts. Pharmaceutically unacceptable salts may nonetheless possess properties such as high crystallinity, which have utility in the practice of the present invention, such as for example utility in process of synthesis, purification or formulation of compounds useful within the methods of the invention.

Suitable pharmaceutically acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid. Examples of inorganic acids include sulfate, hydrogen sulfate, hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric, and phosphoric acids (including hydrogen phosphate and dihydrogen phosphate). Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4- hydroxybenzoic, phenylacetic, mandelic, embonic (or pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, sulfanilic, 2-hydroxyethanesulfonic, trifluoromethanesulfonic, p-toluenesulfonic, cyclohexylaminosulfonic, stearic, alginic, b- hydroxybutyric, salicylic, galactaric, galacturonic acid, glycerophosphonic acids and saccharin (e.g., saccharinate, saccharate). Salts may be comprised of a fraction of one, one or more than one molar equivalent of acid or base with respect to any compound of the invention.

Suitable pharmaceutically acceptable base addition salts of compounds of the invention include, for example, ammonium salts and metallic salts including alkali metal, alkaline earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts made from basic amines such as, for example, N,N’-dibenzylethylene- diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (or N- methylglucamine) and procaine. All of these salts may be prepared from the corresponding compound by reacting, for example, the appropriate acid or base with the compound.

Methods

The invention provides a method of treating or preventing pancreatic cancer, such as but not limited to K-Ras-driven pancreatic cancer, in a subject. In certain embodiments, the method comprises administering to the subject in need thereof a therapeutically effective amount of estrogen and/or a GPCR agonist and a therapeutically effective amount of immunotherapy, such as but not limited to an immune checkpoint inhibitor.

In certain embodiments, the pancreatic cancer comprises pancreatic adenocarcinoma, acinar cell carcinoma, cystadenocarcinoma, pancreatoblastoma, adenosquamous carcinoma, signet ring cell carcinoma, hepatoid carcinoma, colloid carcinoma, undifferentiated carcinoma, undifferentiated carcinoma with osteoclast-like giant cells, solid pseudopapillary tumor, and/or pancreatic mucinous cystic neoplasm.

The invention provides a method of treating or preventing MCC in a subject. In certain embodiments, the method comprises administering to the subject in need thereof a therapeutically effective amount of estrogen and/or a GPCR agonist (such as, for example, a GPER agonist). In other embodiments, the subject is further administered a therapeutically effective amount of immunotherapy, such as but not limited to an immune checkpoint inhibitor.

In certain embodiments, the GPCR is GPER. In other embodiments, the GPER agonist comprises G-l. In certain embodiments, the GPER agonist is selected from the group consisting of estradiol (E2), tamoxifen, fulvestrant, and raloxifene (also known as 6-hydroxy- 2-(4-hydroxyphenyl)-benzothiophen-3-yl]-[4-[2-(l-piperidyl)ethoxy]phenyl]-methanone). In certain embodiments, the estrogen is any substance, natural or synthetic, that mimics the effect of the natural hormone, estrogen. Examples of estrogen contemplated within the invention include, but are not limited to, estrone (El), estradiol (E2), estriol (E3), estetrol (E4), l7P-estradiol, 27-hydroxy cholesterol, dehydroepiandrosterone (DHEA), 7-oxo- DHEA, 7a-hydroxy-DHEA, l6a-hydroxy-DHEA, 7P-hydroxyepiandrosterone, D⁴- androstenedione, A⁵-androstenediol, 3a-androstanediol, 3P-androstanediol, 2-hydroxyestrone, 16-hydroxy estrone, estradiol cypionate, estradiol valerate, estradiol acetate, estradiol benzoate, ethinyl estradiol (EE), mestranol, moxestrol, quinestrol, diethylstilbestrol benzestrol, dienestrol, dienestrol acetate, diethylstilbestrol dipropionate, fosfestrol, hexestrol, methestrol dipropionate, xenoestrogens, phytoestrogens, and/or mycoestrogens.

In certain embodiments, the estrogen and/or GPCR agonist is administered to the subject as a pharmaceutical composition further comprising at least one pharmaceutically acceptable carrier.

Examples of immune checkpoint inhibitors include, but are not limited to, PD-l inhibitors (i.e. Pembrolizumab, Nivolumab, anti-PD-l), PD-L1 inhibitors (i.e. Avelumab, Atezolizumab, anti-PD-Ll), CTLA-4 inhibitors (i.e. Ipilimumab, anti-B7-l/B7-2, anti- CTLA-4), Indoleamine (2, 3)-di oxygenase (IDO 1/2) inhibitors, B7 homolog 3 (B7-H3) inhibitors, lymphocyte activation gene 3 (LAG3) inhibitors, and TIGIT (T cell

immunoreceptor with Ig and ITEM domains) targeting antibodies and agents. In other embodiments, the immune checkpoint inhibitor is co-administered or co-formulated with the estrogen or GPCR agonist.

In certain embodiments, the estrogen or GPCR agonist is administered to the subject by an inhalational, oral, rectal, vaginal, parenteral, topical, transdermal, pulmonary, intranasal, buccal, ophthalmic, intrathecal, intracranial, or intravenous route of

administration. In other embodiments, the estrogen or GPCR agonist is the only anticancer agent administered to the subject. In yet other embodiments, the estrogen or GPCR agonist is the only anticancer agent administered to the subject in an amount sufficient to treat or prevent the cancer in the subject.

In certain embodiments, the subject is a mammal. In yet other embodiments, the mammal is human.

F onmilations/Adni inistration

The relative amounts of the active ingredient, the pharmaceutically acceptable carrier, and any additional ingredients in a pharmaceutical composition of the invention will vary, depending upon the identity, size, and condition of the subject treated. By way of example, the composition may comprise between about 0.005% and about 100% (w/w) of the active agent, or any fractions or multiples thereof.

In certain embodiments, the pharmaceutical compositions useful for practicing the method of the invention may be administered to deliver a dose of between 1 ng/kg/day and 100 mg/kg/day, such as for example 1-50 mg/kg/day. In other embodiments, the

pharmaceutical compositions useful for practicing the invention may be administered to deliver a dose of between 1 ng/kg/day and 1,000 mg/kg/day.

The composition comprising a compound contemplated within the invention can be administered to a mammal as frequently as several times daily, or it may be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every several months or even once a year or less.

It is understood that the amount of compound dosed per day may be administered, in non-limiting examples, every day, every other day, every 2 days, every 3 days, every 4 days, or every 5 days. For example, with every other day administration, a 0.5-5 mg per day dose may be initiated on Monday with a first subsequent 0.5-5 mg per day dose administered on Wednesday, a second subsequent 0.5-5 mg per day dose administered on Friday, and so on. The frequency of the dose will be readily apparent to the skilled artisan and will depend upon any number of factors, such as, but not limited to, the type and severity of the disease being treated, the type and age of the animal, and so forth.

Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions that are suitable for ethical

administration to humans, it will be understood by the skilled artisan that 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 perform such modification with merely ordinary, if any, experimentation. Subjects to which administration of the

pharmaceutical compositions of the invention is contemplated include, but are not limited to, humans and other primates, mammals including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, and dogs.

In certain embodiments, the compositions comprising a compound contemplated within the invention are formulated using one or more pharmaceutically acceptable excipients or carriers. In certain embodiments, the pharmaceutical compositions of the invention comprise a therapeutically effective amount of at least one compound contemplated within the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers, which are useful, include, but are not limited to, glycerol, water, saline, ethanol and other pharmaceutically acceptable salt solutions such as phosphates and salts of organic acids. Examples of these and other pharmaceutically acceptable carriers are described in

Remington’s Pharmaceutical Sciences (1991, Mack Publication Co., New Jersey).

Formulations may be employed in admixtures with conventional excipients, i.e., pharmaceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, intravenous, subcutaneous, enteral, or any other suitable mode of administration, known to the art. In certain embodiments, the administration comprises topical administration. The pharmaceutical preparations may be sterilized and if desired mixed with auxiliary agents, e.g ., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure buffers, coloring, flavoring and/or aromatic substances and the like. They may also be combined where desired with other active agents, e.g. , other analgesic agents.

As used herein,“additional ingredients” include, but are not limited to, one or more of the following: excipients; surface active agents; dispersing agents; inert diluents; granulating and disintegrating agents; binding agents; lubricating agents; sweetening agents; flavoring agents; coloring agents; preservatives; physiologically degradable compositions such as gelatin; aqueous vehicles and solvents; oily vehicles and solvents; suspending agents;

dispersing or wetting agents; emulsifying agents, demulcents; buffers; salts; thickening agents; fillers; emulsifying agents; antioxidants; antibiotics; antifungal agents; stabilizing agents; and pharmaceutically acceptable polymeric or hydrophobic materials. Other “additional ingredients” that may be included in the pharmaceutical compositions of the invention are known in the art and described, for example in Genaro, ed. (1985, Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, PA), which is incorporated herein by reference.

Topical Administration

An obstacle for topical administration of pharmaceuticals is the stratum corneum layer of the epidermis. The stratum corneum is a highly resistant layer comprised of protein, cholesterol, sphingolipids, free fatty acids and various other lipids, and includes cornified and living cells. One of the factors that limit the penetration rate (flux) of a compound through the stratum corneum is the amount of the active substance that can be loaded or applied onto the skin surface. The greater the amount of active substance applied per unit of area of the skin, the greater the concentration gradient between the skin surface and the lower layers of the skin, and in turn the greater the diffusion force of the active substance through the skin. Therefore, a formulation containing a greater concentration of the active substance is more likely to result in penetration of the active substance through the skin, and more of it, and at a more consistent rate, than a formulation having a lesser concentration, all other things being equal.

Formulations suitable for topical administration include, but are not limited to, liquid or semi-liquid preparations such as liniments, lotions, oil-in-water or water-in-oil emulsions such as creams, ointments or pastes, and solutions or suspensions. Such formulations may be applied to the skin directly or through the use of swabs, applicators, spatulas and the like, as well as in the form of transdermal patches. In certain embodiments, the patch minimizes loss of pharmaceuticals through washing, friction, scratching and/or rubbing of the skin. In other embodiments, the patch increases absorption of the pharmaceutical through the skin, while minimizing the exposure of the skin to the pharmaceutical.

Enhancers of permeation may be used. These materials increase the rate of penetration of drugs across the skin. Typical enhancers in the art include ethanol, glycerol monolaurate, PGML (polyethylene glycol monolaurate), dimethylsulfoxide, and the like. Other enhancers include oleic acid, oleyl alcohol, ethoxy diglycol, laurocapram,

alkanecarboxylic acids, dimethylsulfoxide, polar lipids, or N-methyl-2-pyrrolidone. One acceptable vehicle for topical delivery of some of the compositions of the invention may contain liposomes. The composition of the liposomes and their use are known in the art (for example, U.S. Patent No. 6,323,219).

Parenteral Administration

As used herein,“parenteral administration” of a pharmaceutical composition includes any route of administration characterized by physical breaching of a tissue of a subject and administration of the pharmaceutical composition through the breach in the tissue. Parenteral administration thus includes, but is not limited to, administration of a pharmaceutical composition by injection of the composition, by application of the composition through a surgical incision, by application of the composition through a tissue-penetrating non-surgical wound, and the like. In particular, parenteral administration is contemplated to include, but is not limited to, subcutaneous, intravenous, intraperitoneal, intramuscular, intrasternal injection, and kidney dialytic infusion techniques.

Controlled Release Formulations and Drug Delivery Systems

In certain other embodiments, the formulations of the present invention may be, but are not limited to, short-term, rapid-offset, as well as controlled, for example, sustained release, delayed release and pulsatile release formulations.

The term sustained release is used in its conventional sense to refer to a drug formulation that provides for gradual release of a drug over an extended period of time, and that may, although not necessarily, result in substantially constant blood levels of a drug over an extended time period. The period of time may be as long as a month or more and should be a release which is longer that the same amount of agent administered in bolus form.

For sustained release, the compounds may be formulated with a suitable polymer or hydrophobic material that provides sustained release properties to the compounds. As such, the compounds useful within the methods of the invention may be administered in the form of microparticles, for example by injection, or in the form of wafers or discs by implantation.

In one embodiment of the invention, the compounds of the invention are administered to a patient, alone or in combination with another pharmaceutical agent, using a sustained release formulation.

The term delayed release is used herein in its conventional sense to refer to a drug formulation that provides for an initial release of the drug after some delay following drug administration and that may, although not necessarily, includes a delay of from about 10 minutes up to about 12 hours.

The term pulsatile release is used herein in its conventional sense to refer to a drug formulation that provides release of the drug in such a way as to produce pulsed plasma profiles of the drug after drug administration.

The term immediate release is used in its conventional sense to refer to a drug formulation that provides for release of the drug immediately after drug administration.

As used herein, short-term refers to any period of time up to and including about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40 minutes, about 20 minutes, about 10 minutes, or about 1 minute and any or all whole or partial increments thereof after drug administration after drug administration.

As used herein, rapid-offset refers to any period of time up to and including about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40 minutes, about 20 minutes, about 10 minutes, or about 1 minute and any and all whole or partial increments thereof after drug administration.

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents were considered to be within the scope of this invention and covered by the claims appended hereto. For example, it should be understood, that modifications in reaction conditions, including but not limited to reaction times, reaction size/volume, and experimental reagents, such as solvents, catalysts, pressures, atmospheric conditions, e.g ., nitrogen atmosphere, and reducing/oxidizing agents, with art- recognized alternatives and using no more than routine experimentation, are within the scope of the present application.

It is to be understood that wherever values and ranges are provided herein, all values and ranges encompassed by these values and ranges, are meant to be encompassed within the scope of the present invention. Moreover, all values that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the present application.

The following examples further illustrate aspects of the present invention. However, they are in no way a limitation of the teachings or disclosure of the present invention as set forth herein.

EXPERIMENTAL EXAMPLES

The invention is now described with reference to the following Examples. These Examples are provided for the purpose of illustration only, and the invention is not limited to these Examples, but rather encompasses all variations that are evident as a result of the teachings provided herein.

Example 1:

The experiments described herein demonstrate that GPER signaling is tumor suppressive in pancreatic ductal adenocarcinoma (PD AC). 2838c3 PD AC cells were treated with G-l, and a Western blot of the dose response treatment is illustrated in FIG. 1 A. 2838c3 PDAC cells were treated with a pulse or constant 500 nM dose of G-l, and a Western blot of the dose response treatment is illustrated in FIG. 1B. FIG. 1C illustrates a proliferation assay of several PDAC lines treated with 500 nM G-l, * denotes p < 0.05, error bars = ± s.d. FIGs. 1D-1G illustrate Western blots for p-RB, RB, and c-Myc on several PDAC cell lines. FIG. 1H illustrates experimental outline of 2838c3 PD AC -bearing mice treated with vehicle or G- 1, as well as isotype antibody control (2A3) or aPD-l antibody. Treatment was started at day 7 after tumors were palpable n = 5 per group. FIG. II illustrates tumor volumes after G-l treatment. FIG. 1 J illustrates tumor volumes over time of treatment groups. FIG. 1K illustrates a survival curve of mice treated with vehicle or G-l, as well as aPD-l antibody or isotype antibody control (2A3). Significance between groups by the Log-Rank (Mantel-Cox) test is listed in the table in FIG. 1K.

FIG. 2 illustrates proliferation of Merkel cell carcinoma cells in the presence of varying concentrations of G-l .

The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims

CLAIMS What is claimed:

1. A method of treating or preventing pancreatic cancer in a subject, the method comprising administering to the subject in need thereof a therapeutically effective amount of estrogen and/or a selective G protein-coupled estrogen receptor (GPER) agonist, wherein the subject is further co-administered at least one immunotherapeutic agent.

2. The method of claim 1, wherein the pancreatic cancer is K-Ras driven.

3. The method of claim 1, wherein the pancreatic cancer comprises pancreatic adenocarcinoma, acinar cell carcinoma, cystadenocarcinoma, pancreatoblastoma, adenosquamous carcinoma, signet ring cell carcinoma, hepatoid carcinoma, colloid carcinoma, undifferentiated carcinoma, undifferentiated carcinoma with osteoclast-like giant cells, solid pseudopapillary tumor, and/or pancreatic mucinous cystic neoplasm.

4. The method of claim 1, wherein the at least one immunotherapeutic agent comprises an immune checkpoint inhibitor.

5. The method of claim 4, wherein the immune checkpoint inhibitor is selected from the group consisting of a PD-l inhibitor, PD-L1 inhibitor, CTLA-4 inhibitor, TIGIT inhibitor, LAG3 inhibitor, IDO(l/2) inhibitor, and B7-H3 inhibitor.

6. The method of claim 1, wherein the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, are co-administered to the subject.

7. The method of claim 6, wherein the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, are co-formulated.

8. The method of claim 1, wherein administering the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, to the subject allows for greater treatment efficacy than administering the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, separately.

9. The method of claim 1, wherein administering the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, to the subject enhances the anti cancer activity of the estrogen and/or GPER agonist, and the at least one immune checkpoint inhibitor, in the subject.

10. A method of treating or preventing Merkel cell carcinoma (MCC) in a subject, the method comprising administering to the subject in need thereof a therapeutically effective amount of estrogen and/or a GPER agonist.

11. The method of claim 10, wherein the estrogen and/or GPER agonist is/are the only anticancer agent administered to the subject.

12. The method of claim 10, wherein the estrogen and/or GPER agonist are/is the only anticancer agent administered to the subject in an amount sufficient to treat or prevent the MCC in the subject.

13. The method of claim 10, wherein the subject is further co-administered at least one immunotherapeutic agent.

14. The method of claim 13, wherein the immunotherapeutic agent comprises an immune checkpoint inhibitor.

15. The method of claim 14, wherein the immune checkpoint inhibitor is selected from the group consisting of a PD-l inhibitor, PD-L1 inhibitor, CTLA-4 inhibitor, TIGIT inhibitor, LAG3 inhibitor, IDO(l/2) inhibitor, and B7-H3 inhibitor.

16. The method of claim 15, wherein the PD-L1 inhibitor comprises avelumab.

17. The method of claim 13, wherein the estrogen and/or GPER agonist, and the at least one immunotherapeutic agent, are co-administered to the subject.

18. The method of claim 17, wherein the estrogen and/or GPER agonist, and the at least one immunotherapeutic agent, are co-formulated.

19. The method of any one of claims 1-18, wherein the estrogen comprises at least one selected from the group consisting of estrone (El), estradiol (E2), estriol (E3), estetrol (E4), l7P-estradiol, 27-hydroxy cholesterol, dehydroepiandrosterone (DHEA), 7-oxo-DHEA, 7a-hydroxy-DHEA, l6a-hydroxy-DHEA, 7P-hydroxyepiandrosterone, A⁴-androstenedione, A⁵-androstenediol, 3a-androstanediol, 3P-androstanediol, 2-hydroxyestrone, 16- hydroxyestrone, estradiol cypionate, estradiol valerate, estradiol acetate, estradiol benzoate, ethinyl estradiol (EE), mestranol, moxestrol, quinestrol, diethylstilbestrol benzestrol, dienestrol, dienestrol acetate, diethylstilbestrol dipropionate, fosfestrol, hexestrol, methestrol dipropionate, xenoestrogens, phytoestrogens, and mycoestrogens, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof.

20. The method of any of claims 1-18, wherein the GPER agonist comprises at least one selected from the group consisting of G-l, tamoxifen, fulvestrant, and raloxifene, or a salt, solvate, tautomer, enantiomer or diastereoisomer thereof.

21. The method of any of claims 1-18, wherein the GPER agonist comprises at least one selected from the group consisting of:

a molecule of formula

wherein in (I):

bond a is a single or double bond, such that:

if bond a is a double bond, R₃ and R₄ are H, and

if bond a is a single bond, R₃ is selected from the group consisting of H, -OH, -OAc, and halo; R* is selected from the group consisting of H, -OH, -OAc, and -S(o- nitrophenyl); or R₃ and R₄ combine to form a diradical selected from the group consisting of -CH₂-, -0CH₂0-, -0CH(CH₃)0-, and -0C(CH₃)₂0-;

R₅ is selected from the group consisting of H, benzyl, C₃-C₄ alkyl, and acetyl; R₍₅ is selected from the group consisting of H, halo, -N0₂, C₁-C₄ alkyl, -C=CH, -C=C- Si(CH₃)₃ (or -CºC-TMS), -O-benzyl, -OH, -OAc, C₁-C₄ alkoxy, -COOH, and -COO(Ci-C₄ alkyl);

R₇ is selected from the group consisting of H, halo, -N0_2. C₁-C₄ alkyl, -OH, -OAc, and C₁-C₄ alkoxy;

R₈ is selected from the group consisting of H, halo, -N0₂, C₁-C₄ alkyl, -O-benzyl, -N(R)(R), - SR, -COOH, -COO(Ci-C₄ alkyl), -OH, -OAc, C₁-C₄ alkoxy, 3 -thietyl-m ethoxy, - S0₂(morpholino), and -OCH₂CH=CH₂, wherein each occurrence of R is independently selected from the group consisting of H and C₁-C₄ alkyl;

Rio is selected from the group consisting of H, C₁-C₄ alkyl, and halo;

a molecule of formula

, wherein in (II):

Ri is selected from the group consisting of =0 and =N-OH;

R₂ is C₁-C₄ alkyl;

R₅ is selected from the group consisting of H, benzyl and C₁-C₄ alkyl;

22. The method of any of claims 1-18, wherein the GPER agonist comprises at least one selected from the group consisting of:

a molecule of Formula (1-1):

wherein in (1-1):

Ri is selected from the group consisting of =0 and =N-OH;

R₂ is C₁-C₄ alkyl;

bond a is a single or double bond, such that:

if bond a is a double bond, R₃ and R₄ are H, and

R₅ is selected from the group consisting of H, benzyl and C₁-C₄ alkyl;

s is selected from the group consisting of H and halo;

and

a molecule of formula

wherein in (II):

Ri is selected from the group consisting of =0 and =N-OH;

R₂ is C₁-C₄ alkyl;

R₅ is selected from the group consisting of H, benzyl and C₁-C₄ alkyl;

R₇ and R₈ are independently selected from the group consisting of H and C1-C4 alkoxy, or R₇ and R₈ combine to form a diradical selected from the group consisting of -0CH₂0-, - 0CH(CH₃)0- and -0C(CH₃)₂0-,

23. The method of any of claims 1-18, wherein the GPER agonist comprises at least one selected from the group consisting of: G-l;

CMPD1 (rel-l-((3aS,4R,9bR)-4-(benzo[d][l,3]dioxol-5-yl)-3a,4,5,9b-tetrahydro-3H- cyclopenta[c]quinolin-8-yl)ethan-l-one);

CMPD2 (rel-l-((3aS,4R,9bR)-4-(6-bromobenzo[d][l,3]dioxol-5-yl)-5-methyl- 3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-l-one);

CMPD3 (rel-l-((3aS,4R,9bR)-4-(6-bromobenzo[d][l,3]dioxol-5-yl)-2,3,3a,4,5,9b- hexahy dro- lH-cyclopenta[c]quinolin-8-yl)ethan- 1 -one);

CMPD4 (rel-l-((3aS,4R,9bR)-5-benzyl-4-(6-bromobenzo[d][l,3]dioxol-5-yl)- 3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan-l-one);

CMPD5 (rel-l-((3aS,4R,9bR)-4-(2-bromophenyl)-3a,4,5,9b-tetrahydro-3H- cyclopenta[c]quinolin-8-yl)ethan-l-one);

CMPD6 (rel-l-((3aS,4R,9bR)-4-(6-bromobenzo[d][l,3]dioxol-5-yl)-3a,4,5,9b- tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan- 1 -one oxime);

CMPD7 (rel-l-((3aS,4R,9bR)-4-(2-bromo-4,5-dimethoxyphenyl)-3a,4,5,9b- tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan- 1 -one);

CMPD8 (rel-l-((3aS,4R,9bR)-4-(6-chlorobenzo[d][l,3]dioxol-5-yl)-3a,4,5,9b- tetrahydro-3H-cyclopenta[c]quinolin-8-yl)ethan- 1 -one);

CMPD9 (rel-l-((6R,6aS,7aS,l0aR,l0bR)-6-(6-bromobenzo[d][l,3]dioxol-5-yl)-9,9- dimethyl-b^a ^a Oa^Ob-hexahydro-SH-fl^Jdioxolofd^S’^^Jcyclopentafl^-cJquinolin- 2-yl)ethan-l-one);

CMPD 10 (rel- 1 -(( lR,2S,3 aS,4R,9bR)-4-(6-bromobenzo[d] [ 1 ,3 ]dioxol-5-yl)- 1 ,2- dihydroxy-2,3,3a,4,5,9b-hexahydro-lH-cyclopenta[c]quinolin-8-yl)ethan-l-one);

CMPD11 (rel-l-((3aS,4R,9bR)-4-(2-bromo-4,5-dimethoxyphenyl)-2,3,3a,4,5,9b- hexahy dro- lH-cy clopenta[c]quinolin-8-yl)ethan- 1 -one); and

CMPD12 (rel-l-((4S,5aS,6R,l laR)-4,5,5a,6,l l,l la-hexahydro-4,6- methanofl^Jdioxolofd’^’iS^Jbenzofl^-cJacridin-S-y^ethan-l-one),

24. The method of any one of claims 1-23, wherein the subject is further administered at least one additional anticancer treatment.

25. The method of claim 24, wherein the at least one additional anticancer treatment comprises chemotherapy, an engineered chimeric antigen receptor (CAR) T-cell, and/or radiation therapy.

26. The method of claim 25, wherein the chemotherapy is selected from the group consisting of a histone deacetylase inhibitor (HD AC), temozolomide, dacarbazine (DTIC), vemurafenib, dabrafenib and trametinib.

27. The method of any one of claims 1-26, wherein the estrogen or GPER agonist is administered to the subject by at least one administration route selected from the group consisting of inhalational, oral, rectal, vaginal, parenteral, topical, transdermal, pulmonary, intranasal, buccal, ophthalmic, intrathecal, intracranial, and intravenous.

28. The method of any one of claims 1-27, wherein the subject is a mammal.

29. The method of claim 28, wherein the mammal is human.

30. A composition comprising (i) estrogen and/or GPER agonist and (ii) an immune checkpoint inhibitor.

31. The composition of claim 30, wherein the immune checkpoint inhibitor is selected from the group consisting of a PD-l inhibitor, PD-L1 inhibitor, CTLA-4 inhibitor, LAG3 inhibitor, IDO(l/2) inhibitor, TIGIT inhibitor, and B7-H3 inhibitor.

32. A kit comprising (i) estrogen and/or GPER agonist and (ii) an immune checkpoint inhibitor, and instructional material for use thereof in treating or preventing a cancer in a subject.

33. The kit of claim 32, wherein the immune checkpoint inhibitor is selected from the group consisting of a PD-l inhibitor, PD-L1 inhibitor, CTLA-4 inhibitor, LAG3 inhibitor, IDO(l/2) inhibitor, TIGIT inhibitor, and B7-H3 inhibitor.