Enantiomerically pure gper agonists for the treatment of disease states and conditions

Abstract

The present disclosure provides 1) enantiomerically pure compounds SRR G-1 or its derivatives, including specific crystalline forms, salts and co-crystals that modulate G protein-coupled estrogen receptor activity, 2) pharmaceutical and cosmetic compositions comprising enantiomerically pure SRR G-1 or its derivatives and 3) methods of treating or preventing disease states and conditions mediated by these receptors in humans and animals as well as methods for cosmetic conditions and related methods.

Description

[0001] This invention is a divisional application of the patent application filed on July 22, 2019, with application number “201980048925.4” (international application number PCT / US2019 / 042827) and titled “enantiomerically pure GPER agonist for treating disease states and symptoms”.

[0002] Government interests

[0003] This invention was completed with the support of the U.S. government and was granted patent number 2R44CA228695-02 by the National Cancer Institute of the National Institutes of Health. The U.S. government enjoys certain rights to this invention. Summary of the Invention

[0004] Embodiments of the present invention relate to enantiomerically pure agonists of G protein-coupled estrogen receptor (GPER), pharmaceutical compositions comprising enantiomerically pure SRR G-1 or derivatives thereof, and methods for treating disease states and conditions in subjects in need, as well as methods for treating disease states and conditions mediated by GPER receptors.

[0005] Estrogen mediates a variety of complex physiological responses throughout the body. These responses are mediated by the binding of estrogen to its receptors. Classical receptors bind steroids (such as estrogen) and are soluble cytoplasmic / nuclear proteins that act as transcription factors. These receptors are known as estrogen receptors α and β (two closely related proteins) that mediate transcriptional activity. GPER is a 7-transmembrane G protein-coupled receptor that also binds to estrogen with high affinity (Kd~6 nM) and mediates rapid cellular responses, including cyclic adenosine monophosphate signaling, calcium mobilization, and the production of phosphatidylinositol 3,4,5-phosphate.

[0006] Endogenous and / or pharmacological activation of GPER signaling can influence disease development, progression, and / or response to treatment. These diseases include cancers (including cancer prevention, prevention of cancer recurrence, and inhibition of cancer progression; particularly melanoma, pancreatic cancer, lymphoma, uveal melanoma, non-small cell lung cancer, breast cancer, reproductive and other hormone-dependent cancers, leukemia, colon cancer, prostate cancer, and bladder cancer), reproductive disorders (genitourinary tract) (including endometritis, prostatitis, polycystic ovary syndrome, and bladder control), hormone-related disorders, hearing impairment, cardiovascular diseases including hot flashes and excessive sweating, hypertension, stroke, obesity, diabetes, osteoporosis, hematologic disorders, vascular diseases, or many other conditions such as venous thrombosis and atherosclerosis, and central and peripheral nervous system disorders including depression, insomnia, anxiety, neuropathy, multiple sclerosis, neurodegenerative disorders (such as Parkinson's disease and Alzheimer's disease), inflammatory bowel disease, and Crohn's disease. Disease, celiac disease and intestinal-related disorders. Attached Figure Description

[0007] Figure 1 Displays an atomic displacement ellipsoid of the SSR G-1 dichloromethane solvate.

[0008] Figure 2 This shows the packing diagram as viewed along the crystallographic a-axis.

[0009] Figure 3 This shows the packing diagram as viewed along the crystallographic b-axis.

[0010] Figure 4 This shows the packing diagram as viewed along the crystallographic c-axis.

[0011] Figure 5 Display a one-dimensional hydrogen bond network.

[0012] Figure 6 SRR G-1 is shown with marked chiral centers.

[0013] Figure 7 The calculated XRPD plot of the SRR G-1 dichloromethane solvate generated from the single-crystal structure is shown.

[0014] Figure 8 Display the atomic displacement ellipsoid diagram of SRR G-1 A type.

[0015] Figure 9 Showing calculated and experimental XRPD plots of SRR G-1 A type.

[0016] Figure 10 Show XRPD diagrams of SRR G-1 types A, B, and C.

[0017] Figure 11 The thermal analysis diagram of SRR G-1 A type is shown.

[0018] Figure 12 Displays the DVS isotherm of SRR G-1 A type.

[0019] Figure 13 Display the atomic displacement ellipsoid diagram of SRR G-1 B type.

[0020] Figure 14 Showing calculated and experimental XRPD plots of SRR G-1 B type.

[0021] Figure 15 The thermal analysis diagram of SRR G-1 B type is shown.

[0022] Figure 16 The DSC thermal analysis diagram of the SRR G-1 type B and type C mixture is shown.

[0023] Figure 17 Displays the XRPD index results for SRR G-1 C type.

[0024] Figure 18 The thermal analysis diagram of SRR G-1 C type is shown.

[0025] Figure 19 This shows an XRPD overlay plot of the residual solids after the pH solubility test (I / II).

[0026] Figure 20 This shows an XRPD overlay plot of the residual solids after the pH solubility test (II / II).

[0027] Figure 21 This shows the solubility of SRR G-1 free base in biologically relevant media.

[0028] Figure 22 XRPD coverage plot of SRR G-1 after solubility test in SGF.

[0029] Figure 23 XRPD coverage plot of SRR G-1 after solubility test in FaSSIF.

[0030] Figure 24 XRPD coverage plot of SRR G-1 after solubility test in FeSSIF.

[0031] Figure 25 Showing the XRPD plot of SRR G-1 salt.

[0032] Figure 26Display the atomic displacement ellipsoid diagram of SRR G-1 benzenesulfonate type A.

[0033] Figure 27 Showing calculated and experimental XRPD plots of SRR G-1 benzenesulfonate type A.

[0034] Figure 28 The thermal analysis diagram of SRR G-1 benzenesulfonate type A is shown.

[0035] Figure 29 Displays the index results for SRR G-1 camphor sulfonate type A.

[0036] Figure 30 The XRPD plot of SRR G-1 camphor sulfonate type A is shown, from 5 to 19° (2θ).

[0037] Figure 31 The thermal analysis diagram of SRR G-1 camphor sulfonate type A is shown.

[0038] Figure 32 Displays the index results for SRR G-1 naphthalenesulfonate type A.

[0039] Figure 33 The thermal analysis diagram of SRR G-1 naphthalenesulfonate type A is shown.

[0040] Figure 34 Results of a proliferation assay using YUMM1.7 melanoma cells are described. In this assay, cells were treated with either 500 nM of a racemic mixture (G-1) or a single enantiomer of G-1 SRR G-1 and RSS G-1. Dashed lines indicate the initial cell population number. There were n = 5 replicates per group. * indicates p < 0.05, error bar = ±sd

[0041] Figure 35 The plasma concentration of SRR G-1 in rats treated with SRR G-1 free base is shown.

[0042] Figure 36 The plasma concentration of SRR G-1 in rats treated with SRR G-1 benzenesulfonate is shown.

[0043] Figure 37 The plasma concentration of SRR G-1 in rats treated with SRR G-1 naphthalenesulfonate is shown.

[0044] Figure 38 Comparison of plasma concentrations of SRR G-1 in rats treated with SRR G-1 free base, SRR G-1 benzenesulfonate, and SRR G-1 naphthalenesulfonate. Invention Details

[0045] definition

[0046] As used herein, the following terms have the meanings indicated.

[0047] Before describing the compounds, compositions, and methods of the present invention, it should be understood that the invention is not limited to the specific methods, formulations, compounds, compositions, or methods described, as these can vary. It should also be understood that the terminology used in this specification is for the purpose of describing a particular version or embodiment only and is not intended to limit the scope of the embodiments herein, which is limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Although any methods and materials similar to or equivalent to those described herein may be used in the practice or testing of the embodiments herein, preferred methods, apparatus, and materials are described here. All publications mentioned herein are incorporated herein by reference in their entirety. Nothing herein should be construed as an admission that the embodiments herein are not entitled to such prior disclosure due to prior invention.

[0048] It must also be noted that, as used herein and in the appended claims, the singular forms “a”, “an” and “the” include the plural forms unless the context clearly indicates otherwise.

[0049] As used in this article, the term “about” refers to a number that is used with the numerical value plus or minus 20%. Therefore, about 50% means in the range of 40%-60%.

[0050] In embodiments or claims where the term "comprising" is used as a transitional phrase, the embodiment may also be contemplated to replace the term "comprising" with the terms "consisting of" or "substantially composed of".

[0051] As used herein, the terms "consists of" or "consisting of" refer to a compound, composition, formulation, or method that includes only the elements, steps, or components specifically listed in the particular claimed embodiment or claim.

[0052] As used herein, the terms "consisting essentially of" or "consists essentially of" mean that a compound, composition, formulation, or method includes only the elements, steps, or components specifically listed in a particular claimed embodiment or claim, and may optionally include additional elements, steps, or components that do not substantially affect the essential and novel features of that particular embodiment or claim. For example, in a formulation or method for treating a specific condition (e.g., cancer and / or obesity), the only active ingredient is the therapeutic agent specifically listed in the particular embodiment or claim.

[0053] As used herein, the term “its derivatives” refers to any molecular form of the compound to which it is referenced, including but not limited to its salts, pharmaceutically acceptable salts, its esters, its free bases, its solvates, its hydrates, its N-oxides, its clathrates, its prodrugs, its isotopes (e.g., tritium, deuterium), its cocrystals, and any combination thereof.

[0054] The compounds disclosed herein contain asymmetric centers. These centers are indicated by the symbols “R” or “S” depending on the configuration of the substituents surrounding the chiral carbon atom. It should be understood that this invention covers all stereochemical isomers, including diastereomers, enantiomers, and epiomers, as well as mixtures thereof. The individual stereoisomers of the compounds can be prepared by synthesis from commercially available starting materials containing chiral centers, or by preparing mixtures of enantiomeric products followed by separation (e.g., conversion to a mixture of diastereomers), followed by separation or recrystallization, chromatographic techniques (direct separation of enantiomers on a chiral column), or any other suitable method known in the art. Starting compounds having a specific stereochemistry are commercially available or can be prepared and resolved using techniques known in the art. Furthermore, the compounds disclosed herein can exist in geometrically isomeric forms. This invention includes all cis, trans, cis-syn, trans-anti, trans (E), and cis (Z) isomers, and suitable mixtures thereof. Furthermore, the compounds can exist in tautomer forms. This invention provides all tautomers. Additionally, the compounds disclosed herein can exist in non-solvent forms as well as in solvated forms with pharmaceutically acceptable solvents such as water, ethanol, etc. Generally, the solvated forms are considered equivalent to the non-solventized forms.

[0055] As used herein, the terms “chiral purity” and “enantiomer excess” (ee) are interchangeable and can refer to a measurement of the absolute difference between the mole fractions of each enantiomer, and are most often expressed as a percentage. %enantiomer excess can be determined by the following formula:

[0056] %ee=|AB|x100

[0057] Where A and B are the corresponding mole fractions of enantiomers in the mixture, such that A + B = 1. The enantiomer excess of a racemic mixture is 0%, while the enantiomer excess of a single, completely pure enantiomer is 100%. For example, a sample having 70% R isomers and 30% S has an enantiomer excess of 40%. This can also be considered a mixture of 40% pure R and 60% racemic mixture (contributing 30% R and 30% S to the overall composition).

[0058] The term “substantially free” as used alone or in combination herein means that the isomer is not present within the quantitation limits of analytical methods such as nuclear magnetic resonance (NMR), gas chromatography / mass spectrometry (GC / MS), high performance liquid chromatography (HPLC), circular dichroism (CD) or other chemical analytical methods.

[0059] "Pharmaceutically acceptable salts" refer to salts or cocrystals that, within reasonable medical judgment, are suitable for use in contact with patient tissues without excessive toxicity, irritation, allergic reactions, etc., and whose benefits / risks are commensurate with a reasonable ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al., (1977) J. Pharm. Sciences, Vol. 66(1), pp. 1-19, describe in detail representative pharmaceutically acceptable salts. Pharmaceutically acceptable “salt” is any acid addition salt or cocrystal, preferably pharmaceutically acceptable acid addition salt or cocrystal, including but not limited to halide salts such as hydrobromide, hydrochloride, hydrofluoride and hydroiodide; inorganic salts such as nitrates, perchlorates, sulfates and phosphates; organic salts such as sulfonates (methanesulfonic acid, trifluoromethanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid or p-toluenesulfonate), acetic acid, malic acid, fumaric acid, succinic acid, citric acid, benzoic acid, gluconic acid, lactic acid, mandelic acid, mucoic acid, papoic acid, pantothenic acid, oxalic acid and maleate; and amino acid salts such as aspartic acid or glutamate, benzenesulfonate, (+)-(1S)-camphor-10-sulfonate, ethane-1,2-disulfonate, hydrochloric acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, sulfuric acid and p-toluenesulfonate. Pharmaceutically acceptable salts can be monoacid or diacid addition salts, such as dihydrohalic acids, disulfuric acid, diphosphoric acid, or diorganic acid salts. Pharmaceutically acceptable salts are used as chiral or achiral reagents, and their selection does not require any anticipated or known preference based on interaction or precipitation with a specific optical isomer of the product of this disclosure.

[0060] As used herein, the term "therapeuticly acceptable salt" refers to a salt or cocrystalline or zwitterionic form of a compound disclosed herein that is therapeutically acceptable as defined herein and is soluble in water or oil. The salt can be prepared either during the final separation and purification of the compound or by reacting a suitable compound in its free base form with a suitable acid or by replacing the therapeutically acceptable salt with a different salt. Representative acid addition salts include acetates, adipates, alginates, L-ascorbic acid salts, aspartates, benzoates, besylates, hydrogen sulfates, butyrates, camphorates, camphor sulfonates, citrates, digluconates, formates, fumarates, gentianates, glutaric acid, glycerophosphates, glycolates, hemisulfates, heptahydrates, hexanoates, hippurates, hydrochlorides, hydrobromide salts, hydroiodates, 2-hydroxyethanesulfonate (hydroxyethyl sulfonate), lactates, maleates, etc. Malonate, DL-mandelate, mesitylenesulfonate, methanesulfonate, naphthaleneethanesulfonate, nicotinic acid ester, 2-naphthalenesulfonate, oxalate, primate, pectinate, persulfate, 3-phenylpropionate, phosphonate, picrate, trimethylacetate, propionate, pyroglutamic acid, succinate, sulfonate, tartrate, L-tartrate, trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate, p-tosylate, and undecanoate. Similarly, the basic groups in the compounds disclosed herein can be quaternized by methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl, dibutyl, and dipentyl sulfates; decyl, lauryl, myristyl, and steroid chlorides, bromides, and iodides; and benzyl and phenethyl bromides. Examples of acids that can be used to form therapeutically acceptable addition salts include inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid, as well as organic acids such as oxalic acid, maleic acid, succinic acid, and citric acid. Therefore, this invention covers sodium, potassium, magnesium, and calcium salts of the compounds disclosed herein.

[0061] As used herein, the terms "patient" and "subject" are interchangeable and can be considered to refer to any living organism that can be treated with the compounds of the present invention. Thus, the terms "patient" and "subject" can include, but are not limited to, any non-human mammal, primate, or human. In some embodiments, the "patient" or "subject" is an adult, child, infant, or fetus. In some embodiments, the "patient" or "subject" is a human. In some embodiments, the "patient" or "subject" is a mammal, such as a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, primate, or human.

[0062] The terms “therapeutic effective amount” or “therapeutic dose” as used herein are interchangeable and may refer to the amount of an active agent or pharmaceutical compound or composition that elicits a clinical, biological, or medical response in an tissue, system, animal, individual, or human being sought by a researcher, veterinarian, physician, or other clinical professional. A clinical, biological, or medical response may include, for example, one or more of the following: (1) prevention of a disease, condition, or disorder in an individual who may be susceptible to such a disease, condition, or disorder but has not yet experienced or exhibited the pathology or symptoms of such a disease, condition, or disorder; (2) inhibition of a disease, condition, or disorder or prevention of further development of the pathology and / or symptoms of such a disease, condition, or disorder in an individual who is experiencing or exhibiting the pathology or symptoms of such a disease, condition, or disorder; and (3) improvement of a disease, condition, or disorder or reversal of the pathology and / or symptoms experienced or exhibited by an individual who is experiencing or exhibiting the pathology or symptoms of such a disease, condition, or disorder.

[0063] As used herein, the terms “administer,” “administering,” or “administration” refer to the direct administration of a compound or a pharmaceutically acceptable salt or composition of a compound to the subject.

[0064] The term "treatment" can be understood as preventing a specific impairment, disease, or condition, alleviating symptoms associated with a specific impairment, disease, or condition, and / or preventing symptoms associated with a specific impairment, disease, or condition. In some implementations, the term refers to slowing the progression of an impairment, disease, or condition or alleviating symptoms associated with a specific impairment, disease, or condition. In some implementations, the term refers to alleviating symptoms associated with a specific impairment, disease, or condition. In some implementations, the term refers to alleviating symptoms associated with a specific impairment, disease, or condition. In some implementations, the term refers to restoring function impaired or lost due to a specific impairment, disease, or condition.

[0065] The term "prevention" can be understood as preventing a specific obstacle, disease, or condition and / or preventing the recurrence of a specific obstacle, disease, or condition.

[0066] The term "unit dosage form" refers to a physically discrete unit suitable for use as a unit dose in human subjects and other animals, each unit containing a predetermined amount of active substance calculated to produce the desired therapeutic effect, as well as suitable pharmaceutical excipients.

[0067] As used herein, the term “disease” is generally synonymous with and interchangeable with the terms “disorder,” “syndrome,” and “symptom” (as in medical conditions) because they all reflect an abnormal condition of the human or animal body or part thereof that impairs normal functioning, usually manifested as obvious signs and symptoms, and resulting in a reduction in the human or animal’s lifespan or quality of life.

[0068] The term "combination therapy" refers to the administration of two or more therapeutic agents to treat the medical condition or disorder described in this disclosure. Such administration encompasses the combined administration of these therapeutic agents in a substantially simultaneous manner, for example, in the form of a single capsule or dose containing a fixed proportion of the active ingredients, or in the form of multiple separate capsules for each active ingredient. Additionally, such administration also encompasses the sequential use of each type of therapeutic agent in the same patient, with the delivery intervals between the therapeutic agents being 1-24 hours, 1-7 days, or 1 or more weeks. In any case, the treatment regimen will provide the beneficial effect of the combination of drugs in treating the condition or disorder described herein.

[0069] compound

[0070] Many organic compounds exist in an optically active form, meaning they possess the ability to rotate the plane of polarization of light. When describing optically active compounds, the prefixes R and S are used to indicate the absolute configuration of the molecule around its chiral center. For a given chemical structure, these compounds (called stereoisomers) are identical, simply mirror images of each other. Specific stereoisomers can also be called enantiomers, and mixtures of such isomers are often referred to as enantiomers or racemic mixtures.

[0071] Stereochemical purity is important in the pharmaceutical field; eight out of the ten most strictly formulated drugs are chiral. For example, the S-enantiomer of propranolol, a β-adrenergic blocker, is 100 times more effective than the R-enantiomer.

[0072] Embodiments of the present invention cover compounds comprising enantiomerically pure G-1 and methods for treating diseases. G1 is a racemic mixture of the enantiomers 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl) ethyl-1-one (hereinafter referred to as "SRR G-1" or "LNS8801") and 1-((3aR,4S,9bS)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl) ethyl-1-one (hereinafter referred to as "RSS G-1" or "LNS8812").

[0073]

[0074] Enantiomerically pure G1 has been purified to its 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl)ethyl-1-one enantiomer, which is superior to the corresponding 1-((3aR,4S,9bS)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl)ethyl-1-one enantiomer. Unless otherwise specified, SRR G1 or its derivatives include any physical form, including amorphous or any crystalline solid form, such as A, B, C, or combinations thereof.

[0075] In some embodiments, compound 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl) ethyl-1-one (also known as “SRR G-1”) or a derivative thereof has a chiral purity of about 90% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 91% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 92% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 93% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 94% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 95% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 96% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 97% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 97.5% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 98% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.1% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.2% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.3% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.4% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.5% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.6% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.7% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.75% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.8% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.9% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.91% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.92% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.93% or higher.In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.94% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.95% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.96% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.97% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.98% or higher. In some embodiments, SRR G-1 or a derivative thereof has a chiral purity of about 99.99% or higher. In some embodiments, SRR G-1 or a derivative thereof is free of its opposite enantiomer within the limits of quantitation. In some embodiments, SRR G-1 or a derivative thereof is substantially free of its opposite enantiomer.

[0076] In any embodiment of SRR G-1 described herein, the compound is a crystal as demonstrated by XRPD analysis or an amorphous substance or a mixture of crystals and amorphous substances as demonstrated by XRPD analysis.

[0077] In any embodiment of SRR G-1 described herein, the form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)ethyl-1-one is selected from... Figure 10 XRPD diagrams characterizing the A-type crystal form, composed of Figure 10 XRPD images characterizing the B crystal form, composed of Figure 10 The XRPD diagram characterizes the C crystal form, amorphous form, or combination thereof.

[0078] In any embodiment of SRR G-1 described herein, the crystal form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)acetoone is selected from... Figure 10 XRPD diagrams characterizing the A-type crystal form, composed of Figure 10 XRPD images characterizing the B crystal form, composed of Figure 10 XRPD diagrams characterize the C crystal form or combinations thereof.

[0079] In some embodiments, the crystal form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)acet-1-one is determined by... Figure 10 The XRPD diagram characterizes the A-type crystal.

[0080] In any embodiment of SRR G-1 described herein, the form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl) ethyl-1-one is selected from crystal form A, characterized by an XRPD plot having peaks at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86 in degrees 2θ (±0.20); crystal form B, characterized by an XRPD plot having peaks at approximately 13.98, approximately 15.44, approximately 19.67, approximately 21.55, and approximately 22.05 in degrees 2θ (±0.20); and crystal form C, characterized by an XRPD plot having peaks at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86 in degrees 2θ (±0.20). (±0.20) indicates peaks located at approximately 10.73, 12.77, 13.49, 16.09, and 20.60; amorphous; or combinations thereof.

[0081] In any embodiment of SRR G-1 described herein, the crystalline form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)ethyl-1-one is selected from crystalline form A, characterized in that the XRPD plot has peaks at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86, expressed in degrees 2θ (±0.20); crystalline form B, characterized in that the XRPD plot has peaks at approximately 13.98, approximately 15.44, approximately 19.67, approximately 21.55, and approximately 22.05, expressed in degrees 2θ (±0.20); and crystalline form C, characterized in that the XRPD plot has peaks at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86, expressed in degrees 2θ (±0.20). (±0.20) indicates peaks located at approximately 10.73, 12.77, 13.49, 16.09, and 20.60; or combinations thereof.

[0082] In some embodiments, the crystal form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl) ethyl-1-one is crystal form A, characterized in that the XRPD plot has peaks located at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86, expressed in degrees 2θ (±0.20). In some embodiments, the A-form is further characterized by an XRPD plot having peaks at approximately 5.75, approximately 9.56, approximately 10.53, approximately 17.03, approximately 20.54, approximately 20.71, approximately 21.25, approximately 21.86, approximately 24.67, and approximately 28.06, expressed in degrees 2θ (±0.20). In some embodiments, the A-form is further characterized by an XRPD plot having peaks at approximately 5.75, approximately 9.56, approximately 10.53, approximately 10.81, approximately 13.02, approximately 14.66, approximately 14.79, approximately 16.23, approximately 17.03, approximately 20.54, approximately 20.71, approximately 21.25, approximately 21.86, approximately 24.67, and approximately 28.06.

[0083] In some embodiments, the crystal form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)ethyl-1-one is crystal form B, characterized in that the XRPD plot has peaks at approximately 13.98, approximately 15.44, approximately 19.67, approximately 21.55, and approximately 22.05, expressed in degrees 2θ (±0.20). In some embodiments, the B crystal form is further characterized by an XRPD plot having peaks at approximately 13.98, approximately 14.19, approximately 15.44, approximately 19.67, approximately 20.82, approximately 21.55, approximately 22.05, approximately 24.65, approximately 26.18, and approximately 28.18, expressed in degrees 2θ (±0.20). In some embodiments, the B crystal form is further characterized by an XRPD plot having peaks at approximately 7.60, approximately 9.71, approximately 13.98, approximately 14.19, approximately 15.44, approximately 18.61, approximately 19.67, approximately 20.82, approximately 21.55, approximately 22.05, approximately 24.65, approximately 26.18, and approximately 28.18, expressed in degrees 2θ (±0.20).

[0084] In some embodiments, the crystal form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)ethyl-1-one is C-form, characterized in that the XRPD plot has peaks in degrees 2θ (±0.20) at approximately 10.73, approximately 12.77, approximately 13.49, approximately 16.09, and approximately 20.60. In some embodiments, the C crystal form is further characterized by XRPD plots having peaks at degrees 2θ (±0.20) at approximately 7.69, approximately 8.62, approximately 10.73, approximately 12.77, approximately 13.49, approximately 16.09, approximately 19.86, approximately 20.60, approximately 22.05, and approximately 22.98.

[0085] In any embodiment of SRR G-1 or its derivatives described herein, the derivatives are salts or eutectic crystals.

[0086] In any embodiment of SRR G-1 or its derivatives described herein, the derivatives are selected from salts or cocrystals formed with benzenesulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, ethane-1,2-disulfonic acid, hydrochloric acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, sulfuric acid, p-toluenesulfonic acid, or combinations thereof.

[0087] In some embodiments, the derivative thereof is a salt or cocrystal formed with benzenesulfonic acid.

[0088] In some embodiments, the derivative thereof is a salt or cocrystal formed with (+)-(1S)-camphor-10-sulfonic acid.

[0089] In some embodiments, the derivative thereof is a salt or cocrystal formed with naphthalene-2-sulfonic acid.

[0090] In some embodiments, the derivative thereof is a salt or cocrystal formed with benzenesulfonic acid and is characterized by an XRPD plot having peaks in degrees 2θ (±0.20) at the following values: about 4.26, about 6.51, about 6.71, about 16.86, about 18.92, about 19.99, about 20.29, about 20.75, about 21.46, about 22.06, about 22.12, and about 23.99.

[0091] In some embodiments, the derivative thereof is a salt or cocrystal formed with (+)-(1S)-camphor-10-sulfonic acid and is characterized by an XRPD plot having peaks in degrees 2θ (±0.20) at the following values: about 5.97, about 11.98, about 12.69, about 13.41, about 16.23, about 17.79, about 18.03, about 18.77, and about 19.69.

[0092] In some embodiments, the derivative thereof is a salt or cocrystal formed with naphthalene-2-sulfonic acid and is characterized by an XRPD plot having peaks in degrees 2θ (±0.20) at the following values: about 6.17, about 12.63, about 12.84, about 13.75, about 14.39, about 16.79, about 17.07, about 17.64, about 19.22, about 19.44, about 20.43, about 21.26, about 21.78, about 22.60, about 23.38, about 26.07, and about 27.63.

[0093] In any embodiment of SRR G-1 described herein, a concentration of 500 nM of SRR G-1 or its derivatives exhibits approximately 2.5-fold or greater inhibition of cell growth in a YUMM1.7 4-day growth assay compared to racemic mixtures of SRR G-1 and its opposite enantiomers. In some embodiments, SRR G-1 or its derivatives exhibit approximately 3-fold or greater inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit approximately 3.5-fold or greater inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit approximately 4-fold or greater inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit approximately 4.5-fold or greater inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit approximately 5-fold or greater inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit approximately 5.5-fold or greater inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit approximately 6-fold or greater inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit an inhibition of cell growth that is approximately 6.5-fold or greater. In some embodiments, SRR G-1 or its derivatives exhibit an inhibition of cell growth that is approximately 7-fold or greater. In some embodiments, SRR G-1 or its derivatives exhibit an inhibition of cell growth that is approximately 7.5-fold or greater. In some embodiments, SRR G-1 or its derivatives exhibit an inhibition of cell growth that is approximately 8-fold or greater. In some embodiments, SRR G-1 or its derivatives exhibit an inhibition of cell growth that is approximately 8.5-fold or greater. In some embodiments, SRR G-1 or its derivatives exhibit an inhibition of cell growth that is approximately 9-fold or greater. In some embodiments, SRR G-1 or its derivatives exhibit an inhibition of cell growth that is approximately 9.5-fold or greater. In some embodiments, SRR G-1 or its derivatives exhibit an inhibition of cell growth that is approximately 10-fold or greater. Or a range between any two of these values.

[0094] In some embodiments, compounds or derivatives of SRR G1 that are substantially free of their opposite enantiomers and are at a concentration of 500 nM exhibit approximately 7.8-fold or greater inhibition of cell growth in the YUMM1.7 4-day growth assay compared to racemic mixtures of SRR G1 and its opposite enantiomers.

[0095] In any embodiment of SRR G-1 described herein, a concentration of 500 nM of SRR G-1 or its derivative exhibits approximately a 5-fold or greater increase in inhibitory effect on cell growth in a YUMM1.7 4-day growth assay compared to the opposite enantiomer of SRR G-1 or its derivative. In some embodiments, the inhibition of cell growth by SRR G-1 or its derivative exhibits approximately a 10-fold or greater increase. In some embodiments, the inhibition of cell growth by SRR G-1 or its derivative exhibits approximately a 15-fold or greater increase. In some embodiments, the inhibition of cell growth by SRR G-1 or its derivative exhibits approximately a 20-fold or greater increase. In some embodiments, the inhibition of cell growth by SRR G-1 or its derivative exhibits approximately a 25-fold or greater increase. In some embodiments, the inhibition of cell growth by SRR G-1 or its derivative exhibits approximately a 30-fold or greater increase. In some embodiments, the inhibition of cell growth by SRR G-1 or its derivative exhibits approximately a 35-fold or greater increase. In some embodiments, the inhibition of cell growth by SRR G-1 or its derivative exhibits approximately a 40-fold or greater increase. In some embodiments, SRR G-1 or its derivatives exhibit an increase of approximately 45-fold or greater in the inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit an increase of approximately 50-fold or greater in the inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit an increase of approximately 55-fold or greater in the inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit an increase of approximately 60-fold or greater in the inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit an increase of approximately 65-fold or greater in the inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit an increase of approximately 70-fold or greater in the inhibition of cell growth. In some embodiments, SRR G-1 or its derivatives exhibit an increase of approximately 75-fold or greater in the inhibition of cell growth. Or a range between any two of these values.

[0096] In some embodiments, compounds or derivatives of SRR G1 that are substantially free of their opposite enantiomers and are present at a concentration of 500 nM exhibit an increase of approximately 39.5-fold or greater in inhibiting cell growth in the YUMM1.7 4-day growth assay compared to the opposite enantiomers of SRR G1 or their derivatives.

[0097] In any of the embodiments described herein, SRR G-1 or its derivatives have higher desired pharmacological activity compared to RSS G-1 or its derivatives. In any of the embodiments described herein, the presence of RSS G-1 or its derivatives would increase undesirable pharmacological activity in combination therapy with SRR G-1.

[0098] Pharmaceutical Composition

[0099] Some embodiments described herein relate to pharmaceutical or cosmetic compositions comprising an enantiomerically pure SRR G-1 or a derivative thereof of the embodiments described herein and a pharmaceutically or cosmetically acceptable carrier, excipient, or mediator.

[0100] In some embodiments, pharmaceutical or cosmetic compositions containing enantiomerically pure SRR G-1 or its derivatives used according to the embodiments herein may be formulated in a conventional manner using one or more pharmaceutically or cosmetically acceptable carriers or excipients.

[0101] The carrier must be "acceptable," meaning compatible with other components of the formulation and harmless to the recipient. A suitable formulation depends on the chosen route of administration. Any known technology, carrier, and excipient can be used, as understood in the art. The pharmaceutical or cosmetic compositions disclosed herein comprising enantiomerically pure SRR G1 or its derivatives can be manufactured in any manner known in the art, for example, by conventional mixing, dissolving, suspending, granulating, sugar coating, grinding, emulsifying, encapsulating, embedding, or compressing processes.

[0102] Pharmaceutical or cosmetic compositions containing enantiomerically pure SRR G1 or its derivatives include those suitable for oral, rectal, nasal, topical (including skin, buccal, sublingual, and intraocular), vaginal, or parenteral (including intramuscular, subcutaneous, and intravenous) administration. Compositions according to the invention may also be present in the form of bolus, ointment, or paste. Tablets and capsules for oral administration may contain conventional excipients, such as binders, fillers, lubricants, disintegrants, or humectants. The tablets may be coated according to methods well known in the art. Oral liquid formulations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups, or elixirs, or may be present as dry products to be formulated with water or other suitable media prior to use. Such liquid formulations may contain conventional additives, such as suspending agents, emulsifiers, non-aqueous media (possibly including edible oils), or preservatives. When desired, the above formulations may be adapted to provide sustained release characteristics of the active ingredient in the composition using standard methods well known in the art.

[0103] In pharmaceutical compositions comprising enantiomerically pure SRR G-1 or its derivatives according to embodiments of the present invention, it is preferred to formulate the compounds of the present invention by mixing them with a pharmaceutically acceptable carrier. Generally, oral administration of pharmaceutical compositions comprising enantiomerically pure SRR G1 or its derivatives is preferred; however, some pharmaceutical compositions comprising enantiomerically pure SRR G1 or its derivatives may preferably be administered parenterally, particularly in intravenous or intramuscular dosage forms, and via other parenterally routes, such as percutaneous, buccal, subcutaneous, suppository, or other routes, including by inhalation or nasal routes. Oral dosage forms are preferably administered in tablet or capsule form (preferably hard or soft gelatin or other protein or polymer capsules). Intravenous and intramuscular pharmaceutical compositions comprising enantiomerically pure SRR G-1 or its derivatives are preferably administered in sterile saline. Of course, those skilled in the art can modify the formulations within the scope of the teachings of the specification to provide numerous pharmaceutical compositions comprising enantiomerically pure SRR G1 or its derivatives for specific routes of administration without causing instability of the compositions of the present invention or affecting their therapeutic activity.

[0104] Pharmaceutical compositions suitable for parenteral injection containing enantiomerically pure SRR G-1 or its derivatives may comprise physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, or may comprise sterile powders to be reconstituted into sterile injectable solutions or dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents, or carriers include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, etc.), suitable mixtures thereof, triglycerides including vegetable oils (such as olive oil), or injectable organic esters such as ethyl oleate. Appropriate flowability can be maintained, for example, by using a coating such as lecithin, in the case of dispersions by maintaining the desired particle size and / or by using surfactants.

[0105] These pharmaceutical or cosmetic compositions containing enantiomerically pure SRR G-1 or its derivatives may also contain excipients such as preservatives, humectants, emulsifiers, and / or dispersants. Microbial contamination of the composition can be prevented by adding various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc. Isotonic agents, such as sugars, sodium chloride, etc., may also be preferably included. Prolonged absorption of injectable pharmaceutical or cosmetic compositions containing enantiomerically pure SRR G-1 or its derivatives can be achieved by using agents capable of delaying absorption, such as aluminum monostearate and / or gelatin.

[0106] Solid dosage forms of pharmaceutical compositions comprising enantiomerically pure SRR G1 or its derivatives for oral administration include capsules, tablets, powders, granules, stabilization in polymeric glass, dissolution in lipid-based liquids, dissolution in solidified liquids, and dissolution in self-emulsifying lipids. In such solid dosage forms, the active compound is combined with at least one inert, conventional excipient (or carrier) (e.g., sodium citrate or dicalcium phosphate) or (a) a filler or extender (e.g., starch, lactose, sucrose, mannitol, or silicate); (b) a binder (e.g., carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, or acacia); (c) a humectant (e.g., glycerin); (d) a disintegrant (e.g., agar, calcium carbonate, potato or cassava starch, alginate, certain complex silicates, or sodium carbonate); (e) a solution retarder (e.g., paraffin); (f) an absorption enhancer (e.g., quaternary ammonium compounds); (g) a wetting agent (e.g., cetyl alcohol or glyceryl monostearate); (h) an adsorbent (e.g., kaolin or bentonite); and / or (i) a lubricant (e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof). For capsules and tablets, the dosage forms may also contain a buffer.

[0107] Solid pharmaceutical compositions containing enantiomerically pure SRR G1 or its derivatives of similar types can also use excipients such as lactose or milk sugar and high molecular weight polyethylene glycol as fillers for soft-filled or hard-filled gelatin capsules.

[0108] Solid dosage forms (e.g., tablets, sugar-coated pills, capsules, and granules) of pharmaceutical compositions comprising enantiomerically pure SRR G-1 or its derivatives may be prepared by coating or shelling, such as enteric coating, and other methods known in the art. They may also contain light-blocking agents and may have compositions that release one or more active compounds in a delayed manner. Examples of encapsulation compositions that can be used are polymeric substances and waxes. If suitable, the active compound may also be in microencapsulated form with one or more of the above-described excipients.

[0109] Liquid dosage forms of pharmaceutical compositions comprising enantiomerically pure SRR G-1 or its derivatives for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, the liquid dosage forms may also contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethanol, isopropanol, ethyl formate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol, dimethylformamide, oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame seed oil, glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol, fatty acid esters of sorbitan, or mixtures thereof, etc.

[0110] In addition to this inert diluent, the composition may also contain excipients such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and flavoring agents.

[0111] In addition to active compounds, suspensions may also contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol or dehydrated sorbitol esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, or mixtures of these substances.

[0112] If applicable, a pharmaceutical composition containing enantiomerically pure SRR G1 or its derivatives for rectal or vaginal administration can be prepared by combining an active agent and any other compound, along with a suitable non-irritating excipient or carrier (e.g., cocoa butter, polyethylene glycol, or suppository wax, which are typically solid at room temperature but liquid at body temperature, thus melting and releasing the active substance in the rectal or vaginal cavity).

[0113] Dosage forms of pharmaceutical or cosmetic compositions comprising enantiomerically pure SRR G-1 or its derivatives for topical application include ointments, powders, sprays, inhalers, and drops suitable for ocular, ear, or nasal application. The compound is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, and / or propellants that may be required. Ophthalmic preparations, eye ointments, powders, and solutions are also considered to be within the scope of this invention.

[0114] In some embodiments, the pharmaceutical or cosmetic composition comprising enantiomerically pure SRR G-1 or a derivative thereof, used according to the embodiments herein, further comprises at least one sunscreen agent. In some embodiments, the pharmaceutical composition further comprises at least one sunblock agent. In a more oily embodiment, the pharmaceutical or cosmetic composition comprises a formulated sunscreen or sunblock agent and enantiomerically pure SRR G-1 or a derivative thereof.

[0115] Active compounds, pharmaceutical or cosmetic compositions containing enantiomerically pure SRR G-1 or its derivatives can be effective over a wide dosage range and are generally administered at therapeutically effective amounts. However, it should be understood that the amount of compound or composition actually administered is usually determined by a physician based on relevant circumstances, including the condition to be treated, the chosen route of administration, the compound or composition actually administered, the individual patient's age, weight and response, and the severity of the patient's symptoms.

[0116] In some embodiments, a pharmaceutical or cosmetic composition comprising enantiomerically pure SRR G-1 or a derivative thereof may contain about 0.001% to about 50% of one or more compounds or compositions disclosed herein. In some embodiments, the amount of said one or more compounds or compositions is about 0.001% to about 50%, about 0.001% to about 45%, about 0.001% to about 40%, about 0.001% to about 30%, about 0.001% to about 20%, about 0.001% to about 10%, about 0.001% to about 5%, about 0.01% to about 50%, about 0.01% to about 45%, about 0.01% to about 40%, about 0.01% to about 30%, about... 0.01% to about 20%, about 0.01% to about 10%, about 0.01% to about 5%, about 0.05% to about 50%, about 0.05% to about 45%, about 0.05% to about 40%, about 0.05% to about 30%, about 0.05% to about 20%, about 0.05% to about 10%, about 0.1% to about 50%, about 0.1% to about 45%, about 0.1% to about 40%, about 0.1% to about 30%, about 0.1% to about 20%, about 0.1% About 10%, about 0.1% to about 5%, about 0.5% to about 50%, about 0.5% to about 45%, about 0.5% to about 40%, about 0.5% to about 30%, about 0.5% to about 20%, about 0.5% to about 10%, about 0.5% to about 5%, about 1% to about 50%, about 1% to about 45%, about 1% to about 40%, about 1% to about 35%, about 1% to about 30%, about 1% to about 25%, about 1% to about 20%, about 1% to about 15%, about 1 Values ​​within the ranges of approximately 1% to 10%, approximately 1% to approximately 5%, approximately 5% to approximately 45%, approximately 5% to approximately 40%, approximately 5% to approximately 35%, approximately 5% to approximately 30%, approximately 5% to approximately 25%, approximately 5% to approximately 20%, approximately 5% to approximately 15%, approximately 5% to approximately 10%, approximately 10% to approximately 45%, approximately 10% to approximately 40%, approximately 10% to approximately 35%, approximately 10% to approximately 30%, approximately 10% to approximately 25%, approximately 10% to approximately 20%, approximately 10% to approximately 15%, or values ​​within these ranges. Specific examples may include approximately 0.001%, approximately 0.01%, approximately 0.05%, approximately 0.1%, approximately 0.25%, approximately 0.5%, approximately 0.75%, approximately 1%, approximately 5%, approximately 10%, approximately 15%, approximately 20%, approximately 25%, approximately 30%, approximately 35%, approximately 40%, approximately 45%, approximately 50%, approximately 60%, approximately 70%, approximately 80%, approximately 90%, or any two of these values. All of the foregoing terms represent weight percentages of the composition. In some embodiments, the composition is suitable for topical application. In some embodiments, the composition is suitable for oral application.In some embodiments, the composition is suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous, intra-articular, and intramedullary), intraperitoneal, transmucosal, transdermal, rectal, intranasal, local (including skin, buccal, sublingual, and intraocular) or vaginal administration.

[0117] In some embodiments, a compound, pharmaceutical, or cosmetic composition comprising enantiomerically pure SRR G-1 or a derivative thereof is a therapeutically effective amount. In some embodiments, the therapeutically effective dose may be about 0.01 mg to about 1000 mg, about 0.01 mg to about 900 mg, about 0.01 mg to about 800 mg, about 0.01 mg to about 700 mg, about 0.01 mg to about 600 mg, about 0.01 mg to about 500 mg, about 0.01 mg to about 400 mg, about 0.01 mg to about 300 mg, about 0.01 mg to about 200 mg, about 0.01 mg to about 100 mg, 0.1 mg to about 1000 mg, about 0.1 mg to about 900 mg, about 0.1 mg to about 800 mg, about 0.1 mg to about 700 mg, about 0.1 mg to about 600 mg, about 0.1 mg to about 500 mg, about 0.1 mg to about 400 mg, about 0.1 mg to about 300 mg, about 0.1 mg to about 200 mg, about 0.1 mg to about 100 mg, about 1 mg to about 1000 mg, about 1 mg to about 900 mg, about 1 mg to about 800 mg, about 1 mg to about 700 mg, about 1 mg to about 600 mg, about 1 mg to about 500 mg, about 1 mg to about 400 mg, about 1 mg to about 300 mg, about 1 mg to about 200 mg, about 1 mg to about 100 mg, about 10 mg to about 1000 mg, about 50 mg to about 1000 mg, about 100 mg to about 1000 mg, about 200 mg to about 1000 mg, about 300 mg to about 1000 mg, about 400 mg to about 1000 mg, about 500 mg to about 1000 mg, about 10 mg to about 500 mg, about 50 mg to about 500 mg, about 10 mg to about 300 mg, about 50 mg to about 300 mg, from about 100 mg to about 300 mg, about 10 mg to about 150 mg mg, about 50 mg to about 150 mg, about 60 mg to about 120 mg, about 50 mg to about 120 mg, or any two of these values.Specific examples include, for example, any value between the disclosed ranges of about 1000 mg, about 900 mg, about 800 mg, about 700 mg, about 750 mg, about 600 mg, about 500 mg, about 400 mg, about 450 mg, about 300 mg, about 250 mg, about 200 mg, about 175 mg, about 150 mg, about 125 mg, about 120 mg, about 110 mg, about 100 mg, about 90 mg, about 80 mg, about 70 mg, about 60 mg, about 50 mg, about 30 mg, about 20 mg, about 10 mg, about 5 mg, about 1 mg, about 0.1 mg, about 0.01 mg, or more.

[0118] In some embodiments, the therapeutically effective dose may vary depending on factors such as the specific purpose of the treatment, the route of administration of the compound or composition, the patient's health and condition, and the prescribing physician's judgment. The proportion or concentration of a compound or composition in a pharmaceutical or cosmetic composition comprising enantiomerically pure SRRG1 or a derivative thereof can vary depending on a number of factors, including dosage, chemical properties (e.g., hydrophobicity), and route of administration. For example, the compound or composition may be provided in a physiologically buffered aqueous solution comprising about 0.1 to about 10% w / v of the compound or composition for parenteral administration. Some typical dose ranges for the compound or composition are about 1 μg / kg body weight to about 1 g / kg body weight per day. In some embodiments, the dose range is about 0.01 mg / kg body weight to about 100 mg / kg body weight per day. The dose may depend on variables such as the type and extent of disease or disorder progression, the overall health condition of the particular patient, the relative biological efficacy of the selected compound or composition, the formulation of the excipient, and its route of administration. The effective dose can be extrapolated from dose-response curves derived from in vitro or animal model testing systems.

[0119] The amount of a compound, pharmaceutical, or cosmetic composition containing enantiomerically pure SRR G1 or its derivatives administered to a patient will vary depending on the content, purpose of administration (e.g., prevention or treatment), patient condition, method of administration, etc. In therapeutic applications, the composition may be administered to a patient already suffering from the disease in an amount sufficient to cure or at least partially prevent symptoms of the disease and its complications.

[0120] One or more additional therapeutic agents may be present in any pharmaceutical composition comprising enantiomerically pure SRR G-1 or its derivatives described herein.

[0121] The additional therapeutic agents may be selected from, but are not limited to, weight-loss drugs, antihyperglycemic agents, insulin sensitizers, glucagon-like peptide-1 (GLP-1) receptor agonists, sodium-glucose cotransporter 2 (SGLT2) inhibitors, insulin, insulin analogs, sulfonylureas, dipeptidyl peptidase 4 (DPP-4) inhibitors, alpha-glucosidase inhibitors (AGI), bile acid sequestrants (BAS), anti-sympathetic dopamine receptor agonists, incretins, antihypertensive drugs, lipid-modifying agents, anti-obesity agents, immunotherapeutic agents, chemotherapeutic agents, targeted kinase inhibitors, histone deacetylase inhibitors, anti-infective agents, bromodomain inhibitors, and combinations thereof.

[0122] The immunotherapeutic agents may be selected from, but are not limited to, PD-1 inhibitors (pembrolizumab, nivolumab, anti-PD-1), PD-L1 inhibitors (i.e., atezolizumab, acitumab, duvarubicin, anti-PD-L1), CTLA-4 inhibitors (i.e., ipilimumab, anti-B7-1 / B7-2, anti-CTLA-4), IL-2, IL-7, IL-12, oncolytic viruses (Talimogene Laherparepvec), cytosine phosphate-guanosine, oligodeoxynucleotides, and imiquimod. imod), Resiquimod and antibodies targeting T cell immune receptors with Ig and ITIM domains (TIGIT), inducible costimulators (ICOS), lymphocyte activation gene 3 (LAG-3), T-cell immunoglobulin and molecule 3 containing mucin domain (TIM3), T cell activation IG inhibitor containing V domain (VISTA), OX40, glucocorticoid-induced TNF receptor (GITR), CD40, CD47, CD94 / NKG2A, cytotoxic immunoglobulin receptor (KIR) and combinations thereof.

[0123] The chemotherapeutic agents may be selected from, but are not limited to, cyclophosphamide, methotrexate, 5-fluorouracil, doxorubicin, docetaxel, bleomycin, vincristine, dacarbazine, nitrogen mustard, vincristine, procarbazine, etoposide, cisplatin, epirubicin, capecitabine, leucovorin, oxaliplatin, temozolomide, taxanes, and combinations thereof.

[0124] The targeted kinase inhibitor may be selected from, but is not limited to, vemurafenib, dabrafenib, trametinib, vandetanib, SU6656, sunitinib, sorafenib, selumetinib, ruxotinib, pegaptanib, pazopanib, nilotinib, mubritinib, lenvatinib, lapatinib, imatinib, ibrutinib, gefitinib, fostamatinib, erlotinib, erdafitinib, dasatinib, cabozantinib, crizotinib, cobitinib, cetuximab, bosutinib, binimetinib, axitinib, afatinib, adavasertib, and combinations thereof.

[0125] The histone deacetylase inhibitor may be selected from, but is not limited to, vorinostat, romidesin, chidamide, panobinostat, belinostat, valproic acid, givinostat, and combinations thereof.

[0126] The anti-infective agent may be selected from, but is not limited to, Orbactiv, dalvavancin, Sivextro phosphate, clindamycin, linezolid, Bactroban, trimethoprim, sulfamethoxazole, trimethoprim-sulfamethoxazole (Septra or Bactrim), tetracycline, vancomycin, daptomycin, fluoroquinolone, and combinations thereof.

[0127] The bromine domain inhibitor may be selected from, but is not limited to, OTX015 / MK-8628, CPI-0610, BMS-986158, ZEN003694, GSK2820151, GSK525762, INCB054329, INCB057643, ODM-207, RO6870810, BAY1238097, CC-90010, AZD5153, FT-1101, ABBV-744, RVX-000222 and combinations thereof.

[0128] How to use

[0129] This document provides methods for treating or preventing diseases or disorders in subjects in need, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition comprising enantiomerically pure SRR G1 or a derivative thereof according to any embodiment disclosed herein.

[0130] A method for treating or preventing a disease or disorder in a subject in need, comprising administering to the subject a therapeutically effective amount of an enantiomerically pure SRR G1 or a derivative thereof as described herein, wherein treatment with SRR G1 is adjunctive to one or more other therapies selected from surgical therapy, chemotherapy, anti-PD-1 therapy, targeted molecular or antiproliferative therapy or radiofrequency ablation therapy, before, during or after such therapy.

[0131] Some implementations describe a method in which cancers or cells that cause or are involved in the disease or disorder express GPER.

[0132] In any of the embodiments described herein, the subject is a human or an animal.

[0133] In some implementations, the disease or disorder is selected from cancer, endometritis, prostatitis, polycystic ovary syndrome, urinary incontinence, hormone-related disorders, hearing impairment, hot flashes, excessive sweating, hypertension, stroke, ischemia, myocardial infarction, dilated cardiomyopathy, obesity, insulin resistance, osteoporosis, atherosclerosis, menopausal symptoms, inflammation, rheumatoid arthritis, osteoarthritis, lymphoproliferative disorders, myeloproliferative disorders, eosinophilia, histiocytosis, paroxysmal nocturnal hemoglobinuria, systemic mastocytosis, venous thrombosis, embolism, depression, insomnia, anxiety, neuropathy, multiple sclerosis, Parkinson's disease, Alzheimer's disease, inflammatory bowel disease, Crohn's disease, celiac disease, proteinuric nephropathy, vascular disease, and thymic atrophy.

[0134] Some implementations describe methods for preventing or reducing the likelihood of pregnancy after sexual intercourse, which include administering to the subject a therapeutically effective amount of a compound or composition or a derivative thereof according to any implementation disclosed herein.

[0135] Some implementations describe methods for restoring lipid profile in subjects in need, which include administering to the subject a therapeutically effective amount of a compound or composition or a derivative thereof according to any of the implementations disclosed herein.

[0136] Some embodiments describe methods for treating or preventing type 2 diabetes in subjects in need, which include administering to the subject a therapeutically effective amount of a compound or composition or a derivative thereof according to any embodiment disclosed herein.

[0137] Type 2 diabetes is a disease diagnosed by a set of characteristics selected from an A1C level greater than or equal to 6.5%, a fasting plasma glucose (FPG) level greater than 126 mg / dL, and an oral glucose tolerance test (OGTT) level greater than 200 mg / dL. Patients with type 2 diabetes have a higher risk of developing dyslipidemia, hypertension, and atherosclerotic cardiovascular disease (ASCVD). In one embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein symptoms of diabetes are treated. In another embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein the A1C level is reduced to less than 6.5%, between 6.4% and 5.7%, or less than 5.7%. In yet another embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein the fasting plasma glucose (FPG) is reduced to less than 126 mg / dL, between 25 mg / dL and 110 mg / dL, less than 110 mg / dL, or less than 100 mg / dL. In one embodiment, a subject is treated by administration of a compound or composition or a derivative thereof according to any embodiment disclosed herein, wherein an oral glucose tolerance test (OGTT) is reduced to less than 200 mg / dL, between 199 mg / dL and 140 mg / dL, or less than 140 mg / dL. In another embodiment, a subject is treated by administration of a compound or composition or a derivative thereof according to any embodiment disclosed herein, wherein blood pressure is reduced to less than 130 / 80 mmHg, less than 120 / 80 mmHg, less than 110 / 80 mmHg, or less than 100 / 80 mmHg. In yet another embodiment, a subject is treated by administration of a compound or composition or a derivative thereof according to any embodiment disclosed herein, wherein blood glucose levels are reduced to less than 70 mg / dL or less than 50 mg / dL. In the implementation scheme, a subject is treated by administration of a compound or composition or a derivative thereof according to any of the embodiments disclosed herein, wherein the risk of developing dyslipidemia, hypertension or atherosclerotic cardiovascular disease (ASCVD) is reduced by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%.

[0138] The diagnosis of prediabetes is based on a set of characteristics selected from an A1C level of approximately 5.7% to 6.4%, a fasting plasma glucose (FPG) level of approximately 100 mg / dL to 125 mg / dL, and an oral glucose tolerance test (OGTT) level of approximately 140 mg / dL to 200 mg / dL. Subjects with prediabetes may also be diagnosed with impaired glucose tolerance, impaired fasting glucose, or insulin resistance. Subjects with prediabetes have a higher risk of developing hyperglycemia, dyslipidemia, hypertension, atherosclerotic cardiovascular disease (ASCVD), cardiometabolic disease, chronic kidney disease, early-stage retinopathy, retinopathy, cardiovascular disease, and biomechanical complications. In one embodiment, the subject is treated by administration of a compound or composition or a derivative thereof according to any embodiment disclosed herein, wherein the symptoms of prediabetes are treated. In another embodiment, the subject is treated by administration of a compound or composition or a derivative thereof according to any embodiment disclosed herein, wherein the A1C level is reduced to less than 6.4% or less than 5.7%. In one embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein the fasting plasma glucose (FPG) is reduced to less than 125 mg / dL, less than 110 mg / dL, or less than 100 mg / dL. In another embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein the oral glucose tolerance test (OGTT) is reduced to less than 199 mg / dL or less than 140 mg / dL. In yet another embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein blood pressure is reduced to less than 130 / 80 mmHg, less than 120 / 80 mmHg, less than 110 / 80 mmHg, or less than 100 / 80 mmHg. In yet another embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein blood glucose levels are reduced to less than 70 mg / dL or less than 50 mg / dL. In the implementation scheme, a subject is treated by administration of a compound or composition or a derivative thereof according to any of the embodiments disclosed herein, wherein the risk of developing hyperglycemia, dyslipidemia, hypertension, atherosclerotic cardiovascular disease (ASCVD), cardiometabolic disease, chronic kidney disease, early-stage kidney disease, retinopathy, cardiovascular disease, and biomechanical complications is reduced by approximately 5%, approximately 10%, approximately 15%, approximately 20%, approximately 25%, approximately 30%, approximately 35%, approximately 40%, approximately 45%, approximately 50%, approximately 55%, approximately 60%, approximately 65%, approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 95%, or approximately 100%.

[0139] A condition characterized by elevated A1C, glucose, and insulin levels, a homeostasis model of insulin resistance assessment (HOMA-IR), urinary 8-iso-PGF2a, oxidative stress in adipose tissue, and GLUT4 carbonylation can be diagnosed as impaired glucose tolerance, impaired fasting glucose, insulin resistance, prediabetes, or type 2 diabetes. Subjects with the condition described herein are at higher risk of developing prediabetes, type 2 diabetes, hyperglycemia, dyslipidemia, hypertension, atherosclerotic cardiovascular disease (ASCVD), cardiometabolic disease, chronic kidney disease, early-stage retinopathy, retinopathy, cardiovascular disease, and biomechanical complications. In an embodiment, the subject is treated by administration of a compound or composition or a derivative thereof according to any embodiment disclosed herein, wherein the symptoms of the condition are treated. In an embodiment, the subject is treated by administration of a compound or composition or a derivative thereof according to any embodiment disclosed herein, wherein the A1C level is reduced to less than 6.4% or less than 5.7%. In one embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein the fasting plasma glucose (FPG) is reduced to less than 125 mg / dL, less than 110 mg / dL, or less than 100 mg / dL. In another embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein the oral glucose tolerance test (OGTT) is reduced to less than 199 mg / dL or less than 140 mg / dL. In yet another embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein blood pressure is reduced to less than 130 / 80 mmHg, less than 120 / 80 mmHg, less than 110 / 80 mmHg, or less than 100 / 80 mmHg. In yet another embodiment, a subject is treated by administration of a compound or composition or derivative thereof according to any embodiment disclosed herein, wherein blood glucose levels are reduced to less than 70 mg / dL or less than 50 mg / dL. In the implementation scheme, a subject is treated by administration of a compound or composition or a derivative thereof according to any of the embodiments disclosed herein, wherein the risk of developing prediabetes, type 2 diabetes, hyperglycemia, dyslipidemia, hypertension, atherosclerotic cardiovascular disease (ASCVD), cardiometabolic disease, chronic kidney disease, early-stage renal disease, retinopathy, cardiovascular disease, and biomechanical complications is reduced by approximately 5%, approximately 10%, approximately 15%, approximately 20%, approximately 25%, approximately 30%, approximately 35%, approximately 40%, approximately 45%, approximately 50%, approximately 55%, approximately 60%, approximately 65%, approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 95%, or approximately 100%.

[0140] In the implementation scheme, one or more additional therapeutic agents may be selected from weight-loss drugs, antihyperglycemic agents, insulin sensitizers, glucagon-like peptide-1 (GLP-1) receptor agonists, sodium-glucose cotransporter 2 (SGLT2) inhibitors, insulin, insulin analogs, sulfonylureas, dipeptidyl peptidase-4 (DPP-4) inhibitors, alpha-glucosidase inhibitors (AGI), bile acid sequestrants (BAS), antisympathetic dopamine receptor agonists, incretins, antihypertensive drugs, lipid modifiers, and combinations thereof.

[0141] In the implementation scheme, the weight-loss drug is selected from diethylamine acetone, phendimetrazine, phentermine, orlistat, phentermine / topiramate extended-release (ER), lorcaserin, naltrexone ER / bupropion ER, and liraglutide. In the implementation scheme, diethylamine acetone is administered at 25 mg. In the implementation scheme, phendimetrazine is administered at 35 mg or 105 mg. In the implementation scheme, phentermine is administered at 8 mg, 15 mg, 30 mg, or 37.5 mg. In the implementation scheme, orlistat is administered at 60 mg or 120 mg. In the implementation scheme, phentermine / topiramate extended-release is administered at 3.75 mg / day (23 mg), 7.5 mg / day (46 mg), or 15 mg / day (92 mg). In the implementation plan, lorcaserin is administered at 10 mg or 20 mg. In the implementation plan, naltrexone ER / bupropion ER is administered at 8 mg naltrexone / 90 mg bupropion. In the implementation plan, liraglutide is administered at 1.2 mg, 1.8 mg, or 3 mg.

[0142] In the implementation scheme, the hypoglycemic drug is selected from metformin and acarbose. In the implementation scheme, metformin is administered at doses of 500 mg, 625 mg, 750 mg, 850 mg, 2000 mg, 2500 mg, or 1 gram. In the implementation scheme, acarbose is administered at doses of 25 mg, 50 mg, or 100 mg.

[0143] In the embodiments, the insulin sensitizer is selected from thiazolidinedione (TZD), pioglitazone, and rosiglitazone. In the embodiments, pioglitazone is administered at 15 mg, 30 mg, or 45 mg. In the embodiments, rosiglitazone is administered at 2 mg, 4 mg, or 8 mg.

[0144] In the embodiments, the glucagon-like peptide-1 (GLP1) receptor agonist is selected from liraglutide, exenatide, abiglutide, and dulaglutide. In the embodiments, liraglutide is administered at 1.2 mg, 1.8 mg, or 3 mg. In the embodiments, exenatide is administered at 2 mg. In the embodiments, abiglutide is administered at 30 mg or 50 mg. In the embodiments, dulaglutide is administered at 0.75 mg or 1.5 mg.

[0145] In the implementation plan, the sodium-glucose cotransporter 2 (SGLT2) inhibitor is selected from empagliflozin, canagliflozin, and dapagliflozin. In the implementation plan, empagliflozin is administered at doses of 5 mg, 10 mg, 12.5 mg, or 25 mg. In the implementation plan, canagliflozin is administered at doses of 50 mg, 100 mg, 150 mg, or 300 mg.

[0146] In the implementation scheme, the insulin is selected from insulin analogs, basal insulin analogs, neutral protamine zinc insulin (NPH), rapid-acting insulin analogs, and inhaled insulin.

[0147] In the implementation scheme, the insulin analogue is selected from insulin glargine, insulin degludec, and insulin detemir. In the implementation scheme, insulin glargine is administered in doses of 100 units or 300 units. In the implementation scheme, insulin degludec is administered in doses of 30 units, 100 units, 200 units, 300 units, or 600 units. In the implementation scheme, insulin detemir is administered in doses of 100 units or 300 units.

[0148] In the implementation scheme, the rapid-acting insulin analogue is selected from lispro insulin, aspart insulin, and lisglutinin insulin. In the implementation scheme, lispro insulin is administered at doses of 50 units, 75 units, 100 units, 300 units, or 1000 units. In the implementation scheme, aspart insulin is administered at doses of 50 units, 90 units, 210 units, 300 units, 700 units, or 1000 units. In the implementation scheme, lisglutinin insulin is administered at doses of 300 units or 1000 units.

[0149] In the embodiments, the sulfonylurea is selected from acetobenzenesulfonylcyclohexylurea, carbutamide, chlorpropamide, tolhexamide, metahexamide, tolasulfonylurea, tolbutamide, glyburide, glibenclamide, gliclazide, glipizide, gliquidone, glimepiride, and glipizide. In the embodiments, acetobenzenesulfonylcyclohexylurea is administered at 250 mg or 500 mg. In the embodiments, carbutamide is administered at 250 mg or 500 mg. In the embodiments, chlorpropamide is administered at 100 mg or 250 mg. In the embodiments, tolasulfonylurea is administered at 100 mg, 250 mg, or 500 mg. In the embodiments, tolbutamide is administered at 250 mg or 500 mg. In the embodiments, glyburide is administered at 5 mg. In the implementation plan, glipizide is administered at 2.5 mg, 5 mg, or 10 mg. In the implementation plan, glipizide is administered at 1 mg, 2 mg, 3 mg, 4 mg, 6 mg, or 8 mg.

[0150] In the embodiments, the dipeptidyl peptidase 4 (DPP4) inhibitor is selected from linagliptin, saxagliptin, and alogliptin. In the embodiments, linagliptin is administered at doses of 2.5 mg, 5 mg, 10 mg, or 25 mg. In the embodiments, saxagliptin is administered at doses of 2.5 mg or 5 mg. In the embodiments, alogliptin is administered at doses of 6.25 mg, 12.5 mg, or 25 mg.

[0151] In the embodiments, the α-glucosidase inhibitor (AGI) is selected from acarbose, miglitol, and voglibose. In the embodiments, acarbose is administered at 25 mg, 50 mg, or 100 mg. In the embodiments, miglitol is administered at 25 mg, 50 mg, or 100 mg. In the embodiments, voglibose is administered at 0.2 mg or 0.3 mg.

[0152] In the embodiments, the bile acid sequestrant (BAS) is selected from cholestyramine, colestipol, and colesevelam. In the embodiments, cholestyramine is administered at 800 mg, 1 gram, or 4 grams. In the embodiments, colestipol is administered at 1 gram or 5 grams. In the embodiments, colesevelam is administered at 375 mg, 625 mg, 1.875 grams, or 3.75 grams.

[0153] In the implementation scheme, the sympatholytic dopamine receptor agonist is bromocriptine mesylate. In the implementation scheme, bromocriptine mesylate is administered at 0.8 mg, 2.5 mg, or 5 mg.

[0154] In the implementation plan, the hypertension drug is selected from angiotensin-converting enzyme inhibitors (ACEIs), angiotensin II receptor blockers (ARBs), beta-blockers, calcium channel blockers (CCBs), and thiazide diuretics.

[0155] In the embodiments, the lipid-modifying agent is selected from ezetimibe, simvastatin, a monoclonal antibody inhibitor of proprotein convertase subtilisin-kexin type 9 serine protease (PCSK9), evolocumab, alirocumab, fibrates, niacin, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and omega-3 fatty acids. In the embodiments, ezetimibe is administered at 10 mg. In the embodiments, simvastatin is administered at 5 mg, 10 mg, 20 mg, 40 mg, or 80 mg. In the embodiments, evolocumab is administered at 140 mg or 420 mg. In the embodiments, alirocumab is administered at 75 mg, 150 mg, or 300 mg. In the implementation plan, niacin is administered at 375 mg, 500 mg, 750 mg or 1 gram.

[0156] Treatment effectiveness can be assessed by measuring insulin levels in the blood. Normal fasting insulin levels are below 5. A fasting insulin level of approximately 8.0 results in a twice-fold risk of prediabetes, while a fasting insulin level of approximately 25 results in a five-fold risk of prediabetes. In an embodiment, administration to a subject in need of a compound or composition or derivative thereof according to any embodiment disclosed herein may reduce fasting insulin levels to less than 5, less than 8, or less than 25.

[0157] Urinary 8-iso-PGF2a is a recognized marker of oxidative stress-induced lipid peroxidation. Elevated urinary 8-iso-PGF2a levels indicate the development of systemic oxidative stress. Treatment efficacy can be assessed by measuring urinary 8-iso-PGF2a levels in adipose tissue. In practice, administration of a compound or composition or derivative thereof according to any embodiment disclosed herein to a subject in need reduces urinary 8-iso-PGF2a levels.

[0158] Treatment efficacy can be assessed by measuring oxidative stress levels in adipose tissue. Oxidative stress is measured by an increase in any of the following enzymes: superoxide dismutase 2 (SOD2), catalase, glutathione peroxidase, peroxiredoxin, aldehyde dehydrogenase, aldehyde-ketone reductase, and glutathione S-transferase. In embodiments, administration to a subject in need of a compound or composition or derivative thereof according to any embodiment disclosed herein may reduce the levels of one or more of the following enzymes: superoxide dismutase 2 (SOD2), catalase, glutathione peroxidase, peroxiredoxin, aldehyde dehydrogenase, aldehyde-ketone reductase, and glutathione S-transferase.

[0159] Treatment efficacy can be assessed by measuring the carbonylation level of GLUT4. Oxidative stress in overnourished adipose tissue leads to widespread oxidation and carbonylation of numerous proteins, including GLUT4 near glucose transport channels, resulting in loss of GLUT4 activity. Carbonylation and oxidation-induced inactivation of GLUT4 can lead to insulin resistance. In this embodiment, administration to a subject in need of a compound or composition or derivative thereof according to any embodiment disclosed herein reduces the level of GLUT4 carbonylation.

[0160] Some implementation methods describe methods for treating or preventing cancer, preventing cancer recurrence, inhibiting cancer progression, shrinking cancer before further treatment, or reducing circulating tumor cells or metastases in subjects in need, comprising administering to said subject a therapeutically effective amount of a compound or composition or derivative thereof according to any implementation method disclosed herein.

[0161] In some implementations, the cancer is selected from reproductive cancers, hormone-dependent cancers, leukemia, colorectal cancer, prostate cancer, breast cancer, ovarian cancer, endometrial cancer, uterine carcinoma, stomach cancer, rectal cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, cervical cancer, endometrial cancer, ovarian cancer, testicular cancer, bladder cancer, kidney cancer, brain / CNS cancer, head and neck cancer, throat cancer, and Hodgkin's disease. Disease), non-Hodgkin's lymphoma, multiple myeloma, melanoma, acute leukemia, lymphocytic leukemia, hairy cell leukemia, acute myeloid leukemia, Ewing's sarcoma, small cell lung cancer, non-small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, Wilms's tumor, neuroblastoma, oral / pharyngeal cancer, esophageal cancer, laryngeal cancer, kidney cancer, lymphoma, Burkitt lymphoma, sarcoma, angiosarcoma, glioblastoma, medulloblastoma, astrocytoma, and Merkel cell carcinoma.

[0162] In a specific implementation, the cancer is selected from melanoma, colorectal cancer, non-small cell lung cancer, and pancreatic cancer.

[0163] Some implementations describe methods for increasing skin pigmentation or preventing or reversing loss of skin pigmentation in subjects in need, which include administering to said subject a therapeutically effective amount of a compound or composition or a derivative thereof according to any implementation disclosed herein.

[0164] Some implementations describe methods for protecting the skin in subjects in need, which include administering to the subject a therapeutically effective amount of a compound or composition or a derivative thereof according to any of the implementations disclosed herein.

[0165] Some implementations describe methods for protecting the skin (including increasing skin pigmentation) in subjects in need, which include administering to the subject a therapeutically effective amount of a compound or composition or a derivative thereof according to any implementation disclosed herein.

[0166] Some implementations describe methods for protecting the skin from skin cancer in subjects in need, which include administering to the subject a therapeutically effective amount of a compound or composition or a derivative thereof according to any implementation disclosed herein.

[0167] Some implementations describe methods for protecting the skin from skin cancer (including increased skin pigmentation) in subjects in need, which include administering to the subject a therapeutically effective amount of a compound or composition or a derivative thereof according to any implementation disclosed herein.

[0168] In embodiments, the method may include the co-administration of one or more additional therapeutic agents. In embodiments, co-administration may be part of the same pharmaceutical composition described herein comprising enantiomerically pure SRR G-1 or a derivative thereof, or a single pharmaceutical composition comprising enantiomerically pure SRR G-1 or a derivative thereof. In embodiments, co-administration may occur at the same time, substantially at the same time, before, or after the administration of the compositions described herein.

[0169] The other therapeutic agents may be selected from weight-loss drugs, antihyperglycemic agents, insulin sensitizers, glucagon-like peptide-1 (GLP-1) receptor agonists, sodium-glucose cotransporter 2 (SGLT2) inhibitors, insulin, insulin analogs, sulfonylureas, dipeptidyl peptidase 4 (DPP-4) inhibitors, alpha-glucosidase inhibitors (AGI), bile acid sequestrants (BAS), anti-sympathetic dopamine receptor agonists, incretins, antihypertensive drugs, lipid-modifying agents, anti-obesity agents, immunotherapeutic agents, chemotherapeutic agents, targeted kinase inhibitors, histone deacetylase inhibitors, anti-infective agents, bromodomain inhibitors, and combinations thereof.

[0170] The immunotherapeutic agents may be selected from PD-1 inhibitors (pembrolizumab, nivolumab, anti-PD-1), PD-L1 inhibitors (i.e., atezolizumab, acitumab, duvarubicin, anti-PD-L1), CTLA-4 inhibitors (i.e., ipilimumab, anti-B7-1 / B7-2, anti-CTLA-4), IL-2, IL-7, IL-12, oncolytic viruses (Talimogene Laherparepvec), cytosine phosphate-guanosine, oligodeoxynucleotides, imiquimod, and remdesivir. Resiquimod and antibodies targeting T-cell immune receptors with Ig and ITIM domains (TIGIT), inducible costimulators (ICOS), lymphocyte activation gene 3 (LAG-3), T-cell immunoglobulin and molecule 3 containing mucin domain (TIM3), T-cell activation IG inhibitor containing V domain (VISTA), OX40, glucocorticoid-induced TNF receptor (GITR), CD40, CD47, CD94 / NKG2A, cytotoxic immunoglobulin receptor (KIR), and combinations thereof.

[0171] The chemotherapeutic agents may be selected from cyclophosphamide, methotrexate, 5-fluorouracil, doxorubicin, docetaxel, bleomycin, vincristine, dacarbazine, nitrogen mustard, vincristine, procarbazine, etoposide, cisplatin, epirubicin, capecitabine, leucovorin, oxaliplatin, temozolomide, taxanes, and combinations thereof.

[0172] Targeted kinase inhibitors can be selected from vemurafenib, dabrafenib, trametinib, vandetanib, SU6656, sunitinib, sorafenib, selumetinib, ruxotinib, pegaptanib, pazopanib, nilotinib, mubritinib, lenvatinib, lapatinib, imatinib, ibrutinib, gefitinib, fostamatinib, erlotinib, erdafitinib, dasatinib, cabozantinib, crizotinib, cobitinib, cetuximab, bosutinib, binimetinib, axitinib, afatinib, adavasertib, and combinations thereof.

[0173] The histone deacetylase inhibitor may be selected from vorinostat, romidesin, chidamide, panobinostat, belinostat, valproic acid, givinostat, and combinations thereof.

[0174] The anti-infective agent may be selected from, but is not limited to, Orbactiv, dalvavancin, Sivextro phosphate, clindamycin, linezolid, Bactroban, trimethoprim, sulfamethoxazole, trimethoprim-sulfamethoxazole (Septra or Bactrim), tetracycline, vancomycin, daptomycin, fluoroquinolone, and combinations thereof.

[0175] The bromine domain inhibitor may be selected from OTX015 / MK-8628, CPI-0610, BMS-986158, ZEN003694, GSK2820151, GSK525762, INCB054329, INCB057643, ODM-207, RO6870810, BAY1238097, CC-90010, AZD5153, FT-1101, ABBV-744, RVX-000222 and combinations thereof.

[0176] 1. A compound comprising the following formula:

[0177]

[0178] 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl) ethyl-1-one or a derivative thereof, wherein the chiral purity of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl) ethyl-1-one or a derivative thereof is about 90% or higher.

[0179] 2. The compound of claim 1, wherein the compound is a crystal as demonstrated by XRPD analysis or an amorphous substance or a mixture of crystals and amorphous substances as demonstrated by XRPD analysis.

[0180] 3. The compound described in item 1, wherein the chiral purity of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl) ethyl-1-one or a derivative thereof is substantially free of its opposite enantiomers.

[0181] 4. The compound of claim 1, wherein 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl) ethyl-1-one is selected from crystal form A, characterized in that the XRPD plot has peaks at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86, expressed in degrees 2θ (±0.20); and crystal form B, characterized in that the XRPD plot has peaks at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86, expressed in degrees 2θ (±0.20). (±0.20) indicates the following peaks: about 13.98, about 15.44, about 19.67, about 21.55 and about 22.05; C crystal form, characterized in that the XRPD pattern has the following peaks in degree 2θ (±0.20): about 10.73, about 12.77, about 13.49, about 16.09 and about 20.60; amorphous; or combinations thereof.

[0182] 5. The compound of claim 1, wherein the crystal form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)acet-1-one is selected from crystal form A, characterized in that the XRPD plot has peaks located at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86, expressed in degrees 2θ (±0.20); and crystal form B, characterized in that the XRPD plot has peaks located at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86, expressed in degrees 2θ (±0.20). (±0.20) indicates the following peaks: about 13.98, about 15.44, about 19.67, about 21.55 and about 22.05; C crystal form, characterized in that the XRPD pattern has the following peaks, indicated by degree 2θ (±0.20): about 10.73, about 12.77, about 13.49, about 16.09 and about 20.60; or combinations thereof.

[0183] 6. The compound of claim 5, wherein the crystal form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl) ethyl-1-one is crystal form A, characterized in that the XRPD plot has peaks in degrees 2θ (±0.20) at the following locations: about 5.75, about 20.54, about 20.71, about 21.25, and about 21.86.

[0184] 7. The compound described in item 6, wherein the crystalline form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)ethyl-1-one is crystalline form A, which is further characterized by an XRPD plot having peaks in degrees 2θ (±0.20) at the following locations: about 5.75, about 9.56, about 10.53, about 17.03, about 20.54, about 20.71, about 21.25, about 21.86, about 24.67, and about 28.06.

[0185] 8. The compound described in item 7, wherein the crystalline form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)ethyl-1-one is crystalline form A, which is further characterized by an XRPD plot having peaks in degrees 2θ (±0.20) at the following locations: about 5.75, about 9.56, about 10.53, about 10.81, about 13.02, about 14.66, about 14.79, about 16.23, about 17.03, about 20.54, about 20.71, about 21.25, about 21.86, about 24.67, and about 28.06.

[0186] 9. The compound of claim 1, wherein the derivative thereof is a salt or a eutectic.

[0187] 10. The compound of claim 1, wherein the derivative thereof is selected from salts or cocrystals formed with benzenesulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, ethane-1,2-disulfonic acid, hydrochloric acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, sulfuric acid, p-toluenesulfonic acid, or combinations thereof.

[0188] 11. The compound of claim 10, wherein the derivative thereof is a salt or cocrystal formed with benzenesulfonic acid.

[0189] 12. The compound of claim 10, wherein the derivative thereof is a salt or cocrystal formed with (+)-(1S)-camphor-10-sulfonic acid.

[0190] 13. The compound of claim 10, wherein the derivative thereof is a salt or cocrystal formed with naphthalene-2-sulfonic acid.

[0191] 14. A pharmaceutical composition comprising a therapeutically effective amount of the compound described in item 1, and a pharmaceutically acceptable carrier, excipient, or mediator.

[0192] 15. A cosmetic composition comprising a therapeutically effective amount of the compound described in item 1, and a cosmetically acceptable carrier, excipient, or medium.

[0193] 16. A method for treating or preventing a disease or disorder in a subject in need, comprising administering to the subject a therapeutically effective amount of the compound or a derivative thereof described in item 1.

[0194] 17. The method described in item 16, wherein the diseases and disorders are selected from cancer, endometritis, prostatitis, polycystic ovary syndrome, urinary incontinence, hormone-related disorders, hearing impairment, hot flashes, excessive sweating, hypertension, stroke, ischemia, myocardial infarction, dilated cardiomyopathy, obesity, insulin resistance, osteoporosis, atherosclerosis, menopausal symptoms, inflammation, rheumatoid arthritis, osteoarthritis, lymphoproliferative disorders, myeloproliferative disorders, eosinophilia, histiocytosis, paroxysmal nocturnal hemoglobinuria, systemic mastocytosis, venous thrombosis, embolism, depression, insomnia, anxiety, neuropathy, multiple sclerosis, Parkinson's disease, Alzheimer's disease, inflammatory bowel disease, Crohn's disease, celiac disease, proteinuric nephropathy, vascular disease, and thymic atrophy.

[0195] 18. A method for preventing or reducing the likelihood of pregnancy after sexual intercourse, comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of item 14.

[0196] 19. The method of item 16, wherein the subject is a human or an animal.

[0197] 20. A method of treating or preventing type 2 diabetes in a subject in need, comprising administering to the subject a therapeutically effective amount of the compound of claim 1 or a derivative thereof.

[0198] 21. The method described in item 17, wherein the cancer is selected from reproductive cancers, hormone-dependent cancers, leukemia, colorectal cancer, prostate cancer, breast cancer, ovarian cancer, endometrial cancer, uterine carcinosarcoma, stomach cancer, rectal cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, cervical cancer, endometrial cancer, ovarian cancer, testicular cancer, bladder cancer, kidney cancer, brain / CNS cancer, head and neck cancer, pharyngeal cancer, Hodgkin's disease, non-Hodgkin's lymphoma, B-cell lymphoma, T-cell lymphoma, etc. Uveal melanoma, triple-negative breast cancer, multiple myeloma, melanoma, acute leukemia, lymphocytic leukemia, hairy cell leukemia, acute myeloid leukemia, Ewing's sarcoma, small cell lung cancer, non-small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, Wilms' tumor, neuroblastoma, oral / pharyngeal cancer, esophageal cancer, laryngeal cancer, kidney cancer, lymphoma, Burkitt lymphoma, sarcoma, angiosarcoma, glioblastoma, medulloblastoma, astrocytoma, and Merkel cell carcinoma.

[0199] 22. A method for increasing skin pigmentation or preventing or reversing loss of skin pigmentation in subjects in need, comprising administering to said subject a therapeutically effective amount of the compound of item 1 or a derivative thereof.

[0200] 23. A method for protecting the skin in a subject in need, comprising administering to the subject a therapeutically effective amount of the compound of item 1 or a derivative thereof.

[0201] 24. The compound of claim 1 or its derivatives, further comprising one or more additional therapeutic agents selected from anti-obesity agents, immunotherapeutic agents, chemotherapeutic agents, targeted kinase inhibitors, histone deacetylase inhibitors, anti-infective agents, bromine domain inhibitors, and combinations thereof.

[0202] 25. The method of item 17, wherein the cancer or cells causing or involved in the disease or disorder express GPER.

[0203] 26. A method for treating or preventing cancer, preventing cancer recurrence or inhibiting cancer progression in subjects in need, comprising administering to the subject a therapeutically effective amount of the compound of item 1 or a derivative thereof.

[0204] Experimental Section

[0205] Option 1

[0206]

[0207] The synthesis of G-1 is described in Org. Biomol. Chem., 2010, 8, 2252-2259, which is incorporated herein by reference and is described in Scheme 1. A catalytic amount of Sc(OTf)3 (0.492 g, 1.0 mmol) was reacted with anhydrous acetonitrile (2.0 cm⁻¹). 3 The solution in 6-bromopiperaldehyde (2.30 g, 10.0 mmol), p-aminoacetophenone (1.30 g, 10.0 mmol), and cyclopentadiene (3.30 g, 50.0 mmol) were added to acetonitrile (25 cm⁻¹). 3 The mixture was stirred at ambient temperature (~23°C) for 2.0 h. Volatiles were removed under vacuum. The residue was purified by preparative silica gel column chromatography, eluting with ethyl acetate-hexane (10:90) to give G-1 (4.03 g, 98%, dr = 94:6) as a white solid. Minor diastereomers were substantially removed by recrystallization to produce a racemic mixture of SRR G-1 and RSS G-1.

[0208] Example 1: Separation of SRR G-1 and RSS G-1 enantiomers

[0209] Starting with a highly pure G1 sample, (±)1-(4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-(3aS*,4R*,9bR*)-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl)ethyl-1-one (99.4% purity), purchased from Tocris Bioscience, was dissolved in 90:10:0.1 (v / v / v) methyl tert-butyl ether / ethanol / diethylamine and preparative chromatography was performed using a Chrialpak 1A resin column. Elution was performed with 90:10:0.1 (v / v / v) methyl tert-butyl ether / ethanol / diethylamine, and fractions corresponding to each enantiomer were collected and concentrated to solids. Single-crystal X-ray structural analysis identified the first eluted enantiomer as the SRR G-1 enantiomer.

[0210] Example 2: SRR G-1 polymorph screening

[0211] Starting with SRR G-1 prepared according to Example 1, a polymorphism screening study was conducted, analyzing the solids separated from the solid slurry or from the solution through rapid and slow evaporation and cooling (Table 1). Two polymorphs were identified: an anhydrous form designated as type A, and a monodichloromethane solvate designated as type B. At elevated temperatures, type B desolventized to form type C. Amorphous substances were generated from purified SRR G-1 using two different methods: rapid evaporation of an ether solution of SRR G-1 or rotary evaporation from a dichloromethane solution of SRR G-1.

[0212] Table 1

[0213]

[0214]

[0215]

[0216] 1 Unless otherwise stated, all times and temperatures are approximate.

[0217] 2 B = birefringent and NB = non-birefringent when observing the substance using a polarizing microscope.

[0218] Single crystal structure determination of SRR G-1 (Type B)

[0219] Starting with SRR G-1 prepared according to Example 1, suitable single crystals were grown from a dichloromethane solution and analyzed by single-crystal X-ray diffraction. The structure was successfully determined.

[0220] Following the evaporation step, single crystals were formed from the dichloromethane solution. These crystals had a diameter of approximately 0.19 × 0.14 × 0.03 mm. 3 Colorless plates of various sizes are mounted randomly on a polymer ring. This is used in an X-ray tube equipped with a copper anode microfocus seal. Preliminary examination and data collection were performed on a RigakuSuperNova diffractometer with a Decrispilatus3 R 200K hybrid pixel array detector. Using a set angle of 6979 reflections in the range of 3.4920° < θ < 77.1910°, the cell constants and orientation matrix used for data collection were refined by least squares. The space group was determined by the program CrysAlisPro as P212121 (international table no. 19). Data were collected up to a maximum diffraction angle (2θ) of 155.132° at room temperature.

[0221] The frame was integrated with CrysAlisPro. A total of 10,119 reflections were collected, of which 4,368 were unique. Lorentz and polarization corrections were applied to the data. The linear absorption coefficient of Cu Kα radiation was 5.144 mm. -1 Empirical absorption correction was performed using CrysAlisPro. Transmission coefficients ranged from 0.676 to 1.000. Average equivalent reflection intensity was calculated. Based on intensity, the consistency factor for the average value was 3.4%.

[0222] The structure was resolved using a direct method with Shelxt. The remaining atoms were included in a subsequent difference-Fourier synthesis. The structure was refined using Shelxt-2014. The hydrogen atoms on SRR G-1 were refined independently. Dichloromethane hydrogen atoms were included in the refinement but were constrained to ride on the atoms they were bonded to. The structure was refined in full-matrix least squares by minimizing the following function:

[0223]

[0224] The weight w is defined as 1 / [δ] 2 (Fo 2 )+(0.0464P) 2 +(0.1905P)], where P=(F o 2 +2F c 2 ) / 3. The scattering factor is taken from the "International Crystallographic Tables". Of the 4368 reflections used in the refinement, only reflections with an intensity greater than twice their uncertainty [I>2δ(I)] were used to calculate the fitting residual R. The final refinement loop includes 334 variable parameters, 0 constraints, and has converged to the following unweighted and weighted agreement factors:

[0225] 0.0348

[0226] 0.0905

[0227] The observed standard deviation per unit weight (goodness-of-fit) was 1.07. The highest peak electron density in the final difference Fourier transform was 0.398 e / Å. 3 The value of the smallest negative peak is -0.438 e / Å. 3 .

[0228] Calculated X-ray powder diffraction (XRPD) patterns. Calculated XRPD patterns for Cu radiation were generated using Mercury and atomic coordinates, space group, and cell parameters from the single-crystal structure. Atomic displacement ellipsoids and packing diagrams. Atomic displacement ellipsoids were prepared using Mercury. Atoms are represented by anisotropic thermal ellipsoids with a 50% probability. Packing diagrams and additional diagrams were generated using Mercury. Hydrogen bonds are represented as dashed lines. Evaluation of chiral centers using Platon. Absolute configuration was evaluated using the canonical rules of molecular chirality.

[0229] The crystal system is orthorhombic, and the space group is P212121. The cell parameters and calculated volume are: a = 6.43156(10) Å, b = 13.0752(2) Å, c = 25.2941(4) Å, α = 90°, β = 90°, γ = 90°, V = 2127.09(6) Å. 3 The standard uncertainty is expressed in crystallographic bracket notation, for example, 0.123(4) equals 0.123 ± 0.004. The formula weight is 497.20 g mol. -1 With Z=4, the calculated density is 1.553 g / cm³. -3 Table 2 summarizes further details regarding the crystal data and crystallographic data collection parameters. As the fitting residual R is 0.0348 (3.48%), the quality of the obtained structures is very high. R-factors cited in the range of 2%–6% represent the most reliably determined structures.

[0230] Table 2 Crystal data and data collection parameters SRR G-1 (Type B)

[0231]

[0232]

[0233] The atomic shift ellipsoid diagram of the SSR G-1 dichloromethane solvate is shown below. Figure 1As shown. The molecules observed in the asymmetric units of the single-crystal structure are consistent with the molecular structures of the proposed SSR enantiomers. Figure 1 The asymmetric unit shown contains an SSR G-1 molecule and a dichloromethane molecule.

[0234] The packing diagrams observed along the crystallographic axes a, b, and c are shown respectively. Figure 2-4 In the middle, hydrogen bonding between amines and carbonyl groups on adjacent molecules leads to the formation of a one-dimensional hydrogen bond network along the b-axis, such as... Figure 5 As shown.

[0235] The absolute structure of a crystal can be determined by analyzing anomalous X-ray scattering. Anomalous scattering is assessed by the intensity difference between Friedel pairs. For values ​​reaching θ... 最大 The reflectance data, with Friedel coverage of 95%, is used. The precise parameter x (called the Flack parameter) encodes the relative abundance of the two components in the inverted bicrystal. The structure contains the fraction 1-x of the model to be refined and its inverse x. Assuming low standard uncertainty, the Flack parameter should be close to 0 if the solved structure is correct, and close to 1 if the inverted model is correct. Figure 1 The structure of the SSR G-1 dichloromethane solvate shown has a measured Flack parameter of -0.018 and a standard uncertainty of 0.013, indicating that it has a strong inversion-distinguishing power.

[0236] Further information about the absolute structure can be assessed by applying Bayesian statistics to the Bijvoet discrepancies. This analysis provides a range of probabilities for different hypotheses of the absolute structure. The analysis yields the Hoofty parameter, which is interpreted in the same way as the Flack x parameter. Furthermore, this analysis yields the probabilities of three absolute structures: correct, incorrect, or racemic bicrystal. For the current dataset, the (equivalent Flack) Hoofty parameter is -0.020 (11), the probability of the correct structure is 1.000, and the probabilities of the incorrect structure or racemic bicrystal are both less than 10. -200 .

[0237] Absolute configuration such as Figure 6 As shown. This is consistent with the configuration of SRR G-1.

[0238] Figure 7 The calculated XRPD plot of the SRR G-1 dichloromethane solvate generated from the single-crystal structure is shown. The calculated XRPD plot is equivalent to the XRPD plots assigned to a large number of samples identified as exhibiting type B XRPD plots (in the polymorphic screening study outlined in Table 1).

[0239] A table showing the positional parameters and their estimated standard deviations, anisotropic displacement factor coefficients, bond spacing, bond angles, hydrogen bond angles, and torsional angles is provided.

[0240] Table 3 Location parameters and their standard deviations for estimation

[0241]

[0242]

[0243]

[0244] Hydrogen atoms are refined isotropically, except for H22A & B, which are included in the calculation of the structure factor but not refined.

[0245] Table 4 Anisotropic Displacement Factor Coefficients

[0246]

[0247]

[0248] The anisotropic temperature factor is in the form of:

[0249] exp[-2h 2 a *2 U(1,1)+k 2 b *2 U(2,2)+l 2 c *2 U(3,3)+2hka * b * U(1,2)+2hla * c * U(1,3)+2klb * c * U(2,3)]

[0250] Where a * b * and c * It is the reciprocal lattice constant.

[0251] Table 5 Bond distances (Ångströms)

[0252]

[0253] The numbers in parentheses are the estimated standard deviation of the least significant figures.

[0254] Table 6 Key Angles

[0255]

[0256]

[0257] The numbers in parentheses are the estimated standard deviation of the least significant figures.

[0258] Table 7 Hydrogen bond distances (unit: angstroms) and angles (degrees)

[0259]

[0260] The numbers in parentheses are the estimated standard deviation of the least significant figures.

[0261] Table 8. Torsion Angle (degrees)

[0262]

[0263]

[0264] The numbers in parentheses are the estimated standard deviation of the least significant figures.

[0265] Starting with SRR G-1 prepared according to Example 1, single crystals of suitable crystal form A were grown from isopropanol solution and analyzed by single-crystal X-ray diffraction. The structure was successfully determined. The crystals had dimensions of approximately 0.203 × 0.137 × 0.033 mm. 3 Single-crystal X-ray analysis was performed on a colorless plate, which was randomly oriented and mounted on a polymer ring. This was done in an X-ray tube equipped with a copper anode microfocus sealed X-ray tube. Preliminary examination and data collection were performed on a Rigaku SuperNova diffractometer with a Decrispilatus3 R 200K hybrid pixel array detector. The cell constants and orientation matrix used for data collection were refined by least squares with a set angle of 9009 reflections in the range of 4.7640° < θ < 77.3860°. The space group was determined to be P212121 by the program CrysAlisPro. Data was collected up to a maximum diffraction angle (2θ) of 155.264 at room temperature.

[0266] The frame was integrated with CrysAlisPro. A total of 17,299 reflections were collected, of which 7,561 were unique. Lorentz and polarization corrections were applied to the data. The linear absorption coefficient of Cu Kα radiation was 3.206 mm. -1 Empirical absorption correction was performed using CrysAlisPro. Transmission coefficients ranged from 0.733 to 1.000. Average equivalent reflection intensity. Based on intensity, the consistency factor for the average value was 2.74%.

[0267] The structure was resolved using a direct method with Shelxt. The remaining atoms were determined in a subsequent difference Fourier synthesis. The structure was refined using Shelxt-2014. Hydrogen atoms were refined independently. The structure was refined in full-matrix least squares by minimizing the following function:

[0268]

[0269] The weight w is defined as 1 / [δ] 2 (F o 2 )+(0.0401P) 2 +(0.3205P)], where P=(F o 2 +2F c 2 ) / 3. The scattering factor is taken from the "International Crystallographic Tables". Of the 7561 reflections used in the refinement, only reflections with an intensity greater than twice their uncertainty [I>2δ(I)] were used to calculate the fitting residual R. The final refinement loop includes 613 variable parameters, 0 constraints, and has converged to the following unweighted and weighted agreement factors:

[0270] 0.0325

[0271] 0.0813

[0272] The observed standard deviation per unit weight (goodness-of-fit) was 1.04. The highest peak electron density in the final difference Fourier transform was 0.319 e / Å. 3 The value of the smallest negative peak is -0.454 e / Å. 3 .

[0273] The crystal system is orthorhombic, and the space group is P212121. The cell parameters and calculated volume are: a = 6.50106(9) Å, b = 17.3547(2) Å, c = 32.6957(4) Å, α = 90°, β = 90°, γ = 90°, V = 3688.85(9) Å 3 The molecular weight is 412.27 g mol. -1 With Z=8, the calculated density is 1.485 g / cm³. -3 Table 9 summarizes further details regarding the crystal data and crystallographic data collection parameters. The atomic displacement ellipsoid of type A is shown below. Figure 8As shown. The asymmetric unit shown contains two enantiomeric SRRG-1 molecules. Structurally, the absolute structure was finally determined. SRRG-1 contains three chiral centers located at C114 (C214), C113 (C213), and C19 (C29), which are bonded in R, S, and R configurations, respectively. Figure 9 The paper provides the calculated XRPD plot of the A-type generated from the single crystal structure and compares it with the experimental plot.

[0274] Table 9 Crystal data and data collection parameters for SRR G-1 A type

[0275]

[0276]

[0277] Example 3: Crystal form and polymorph data

[0278] SRR G-1 forms two distinct polymorphs, type A and type C; a solvate, type B; and an amorphous substance. Figure 10 The XRPD diagrams of the crystal forms are compared. Form B is a single DCM solvate that desolvates to form C upon exposure to high temperatures of 100–120 °C. Form C, the desolvate, exhibits melting initiation around 129 °C. Form A is thermodynamically stable at all temperatures (unidirectionally correlated with form C) and exhibits melting initiation around 178 °C. Amorphous substances are physically unstable and crystallize to form A upon exposure to high temperatures or high humidity. These forms will be discussed in more detail in subsequent sections below.

[0279] Crystal type A

[0280] Form A is anhydrous, with a melting point close to 178°C. Form A is the most thermodynamically stable form and is unidirectionally correlated with form C. Form A is typically prepared using various crystallization techniques employing a variety of organic solvents and organic / aqueous solvent systems other than dichloromethane.

[0281] Table 10 lists the observed XRPD peaks for crystal form A.

[0282] Table 10

[0283]

[0284]

[0285]

[0286] The thermal analysis diagram of type A is as follows: Figure 11As shown. Thermogravimetric analysis (TGA) data showed no weight loss up to 266°C, consistent with the anhydrous form. DSC showed a single endothermic reaction with an onset temperature close to 176°C (68 J / g). The event was visually confirmed as a melt on the heating plate. Discoloration was observed during melting, likely due to decomposition.

[0287] The dynamic vapor adsorption isotherm of type A indicates that this crystal form exhibits low hygroscopicity. Figure 12 The weight change during the adsorption / desorption cycle was less than 0.3% by weight. No hysteresis was observed. The substance recovered from the dynamic vapor adsorption experiment was type A (via XRPD).

[0288] B crystal form

[0289] Type B is a monodichloromethane solvate, typically generated from DCM as a mixture with Type C (the desolvated form). When exposed to temperatures between 100 and 120°C, Type B will completely desolvate to form Type C.

[0290] The single-crystal structure of type B is known. The crystal system is orthorhombic, and the space group is P212121. The cell parameters and calculated volume are: a = 6.43156(10) Å, b = 13.0752(2) Å, c = 25.2941(4) Å, α = 90°, β = 90°, γ = 90°, V = 2127.09(6) Å. 3 The formula weight is 497.20 g mol. -1 With Z=4, the calculated density is 1.553 g / cm³. -3 The asymmetric unit comprises an enantiopure SRR G-1 molecule and a dichloromethane molecule. The structure contains molecules located at C8, C9, and C13 (see [link to relevant documentation]). Figure 13 The three chiral centers of the single-crystal structure are bonded in R, S, and R configurations, respectively. The calculated XRPD plot of the B-type crystal produced by the single-crystal structure is shown in... Figure 14 The data is provided in the documentation and compared with experimental data.

[0291] Table 11 lists the observed XRPD peaks of crystal form B.

[0292] Table 11

[0293]

[0294]

[0295] The thermal analysis diagram of type B is as follows: Figure 15As shown. Thermogravimetric analysis (TGA) curves show that at temperatures up to 177°C, the weight loss is approximately 15.3%, consistent with the volatilization of 0.9 mol / mol DCM. This loss occurs simultaneously with the endothermic desolvation (up to 104°C) and exothermic recrystallization (up to 140°C) shown in the DSC thermal analysis curve. The recrystallized material exhibits an endothermic final melting, with an onset temperature close to 176°C, consistent with the melting of type A. Figure 16 The DSC thermal analysis plots of the mixture of types B and C are provided. The mixture exhibits endothermic desolvation (up to 101 °C), followed by endothermic melting in the type C desolvation form (starting around 128 °C).

[0296] The physical stability of type B was investigated. Exposure to 120°C resulted in complete desolvation to form type C. Exposure to 90–100°C for 25 minutes resulted in almost complete desolvation. Vacuum at 70°C (or below) was insufficient for desolvation.

[0297] C-crystal form

[0298] Type C is a desolvent with a melting start temperature close to 129°C, generated by desolventizing Type B (single DCM solvate).

[0299] The XRPD pattern of type C has been successfully indexed, indicating that it consists of a single-crystal phase. Figure 17 Assuming the chemical composition is correct, it has an orthorhombic unit cell containing four SRR G-1 molecules. Therefore, the value is calculated to be 462.88 Å based on the index results. 3 The formula per unit volume will be consistent with the anhydrous form, and its calculated density is 1.479 g / cm³. -3 .

[0300] Table 12 lists the observed XRPD peaks for the C crystal form.

[0301] Table 12

[0302]

[0303]

[0304] The differential scanning calorimetry (DSC) thermal analysis curve of type C is shown below. Figure 18 As shown, DSC exhibits a single endothermic reaction, with an initial temperature close to 129°C (23 J / g). This event was visually confirmed as a melt on the heating plate.

[0305] amorphous

[0306] The physical stability of the amorphous material produced from purified SRR G-1 was investigated. The amorphous material exposed to high temperature (60°C for 4 days) or high humidity (75% RH for 12 days) crystallized into form A. This indicates that the amorphous material is unstable under the evaluated conditions.

[0307] Relative thermodynamic stability of crystal form

[0308] Phase transitions in solids can be thermodynamically reversible or irreversible. A crystalline form that undergoes a reversible phase transition at a specific transition temperature is called an enantiotropic polymorph. If the crystalline forms cannot interconvert under these conditions, the system is unidirectional (a thermodynamically stable form). Several rules help predict the relative thermodynamic stability of polymorphs and whether the relationship between polymorphs is enantiotropic or monotropic. Density and heat, rationally fused based on statistical mechanics, are used here to guide the determination of unidirectionality or enantiotropicity.

[0309] The density rule is based on the Kitaǐgorodskiǐ principle of closest packing of molecular crystals, which states that for non-hydrogen-bonded systems at absolute zero, the most stable polymorph will have the highest density due to stronger intermolecular van der Waals interactions. Therefore, according to this rule, the crystal structure with the most efficient packing will also have the lowest free energy. It is assumed that hydrogen bonding (the long-distance effect) is not a major parameter of crystal packing. The densities determined from the indexing results of the single-crystal structure of type A and type C show that type A is more stable than type C at absolute zero (1.485 and 1.479 g cm⁻¹, respectively). -3 ).

[0310] The melting initiation and heat of fusion obtained from calorimetric data can be used to estimate the relative physical stability of crystal forms at all temperatures. Figure 11 and 18 According to the heat of fusion rule, if the form with the higher melting point has a lower heat of fusion, the two forms are interconvertible; otherwise, they are unidirectional. The density and heat of fusion rules of this system seem to align with this unidirectional relationship.

[0311] Interconversion experiments were conducted to test the thermodynamic relationship between polymorphs A and C. Interconversion, or competitive slurry experiments, are solution-mediated processes that provide a growth pathway for the less soluble (more stable) polymorph at the expense of the more soluble polymorph. The more stable polymorph obtained from interconversion experiments is independent of the solvent used, except for solvation or degradation, because the thermodynamically more stable polymorph has lower energy and therefore lower solubility. The choice of solvent affects the kinetics of polymorph conversion, but not the thermodynamic relationship between the polymorphs. A saturated solution was produced and then added to a mixture consisting of approximately equal amounts of the two polymorphs. The sample was slurried for nine days, and the solids were collected and analyzed by XRPD. The results of the interconversion studies confirmed that polymorph A is thermodynamically more stable than polymorph C at room temperature. The experimentally determined relative stability at room temperature, the relative stability suggested by the density rule at absolute zero, and the unidirectionality determined by the heat of fusion rule all suggest that polymorph A is more stable than polymorph C at any temperature.

[0312] Solubility of SRR G-1 Type A

[0313] Table 13 Approximate solubility of purified SRR G-1 type A

[0314]

[0315] Solubility measurement of SRR G-1 (Type A) in pH buffer solution

[0316] The solubility of SRR G-1 (A crystal form) was measured over 24 hours in pH buffer (2.0–8.0) at 37°C. The results are summarized in Table 14. XRPD plots are shown below. Figure 19 and Figure 20 As shown. No change in the form of the compound was observed in any pH buffer after 24 hours. The solubility of SRR G-1 in the pH range of 2.0–8.0 is less than 0.72 μg / mL.

[0317] Table 14 Summary of solubility measurements of SRR G-1 (Type A) in pH buffer solution

[0318]

[0319] Solubility Measurement of SRR G-1 (Type A) in Biologically Relevant Media

[0320] The kinetic solubility of SRR G1 (A crystal form) was measured at 37 °C in three biorelevant media (SGF (pH 1.8), FaSSIF (pH 6.5), and FeSSIF (pH 5.0)) for 1, 2, 4, and 24 hours. The results are summarized in Table 15 and... Figure 21 The XRPD plot of the wet pancake is shown in the image. Figure 22-24 No changes in sample form were observed in the three biorelevant media at 1, 2, 4, and 24 hours, respectively. The highest solubility of SRR G-1 (~0.037 mg / mL) was observed in FeSSIF.

[0321] Table 15 Summary of solubility measurements in biologically relevant media

[0322]

[0323] The pKa and LogD of compound SRR G-1 were predicted using MarvinSketch 5.6.0.2. 7.4 The results showed that the pKa of SRR G-1 was 1.90 (base, pH range 0-14), and the LogD... 7.4 The pKa values ​​were 5.32 and LogP was 5.32. pKa titration showed no observed pKa values ​​in the range of 3–11, consistent with the predicted results. LogD was measured using a shake-flask method with a pH 7.4 phosphate buffer and n-octanol solvent system. 7.4 Table 16 shows the LogD 7.4 Detailed results for LogP. Given the low solubility of the free SRR G-1 base in aqueous phase (<0.82 μg / mL) and the small pKa value, LogD... 7.4 The value was determined to be >3.22 and LogP was also >3.22.

[0324] Table 16 LogD of compound SRR G-1 7.4 and LogP

[0325]

[0326] *LogP=LogD (pH) +Log[1+10 (pKa-pH) ]

[0327] Instrumentation Technology

[0328] Differential scanning calorimetry (DSC)

[0329] DSC was performed using a Mettler-Toledo DSC3+ or DSC822e differential scanning calorimeter. Tau lag was adjusted using indium, tin, and zinc. Temperature and enthalpy were adjusted using octane, phenyl salicylate, indium, tin, and zinc. This adjustment was then verified using octane, phenyl salicylate, indium, tin, and zinc. The sample was placed in a sealed aluminum DSC pot, and the weight was accurately recorded. The pot was then inserted into the DSC cell. A weighing aluminum pot configured as the sample pot was placed on the reference side of the cell. The pot lid was punctured before sample analysis. The sample was analyzed from -30°C to 250°C at a rate of 10 °C / min. Cyclic DSC was performed by heating from -30°C to 100°C, returning to -30°C, and then heating to 250°C at a rate of 10 °C / min.

[0330] Dynamic vapor adsorption / desorption (DVS)

[0331] Moisture adsorption / desorption data were collected on a VTI SGA-100 vapor adsorption analyzer. NaCl and PVP were used as calibration standards. Samples were not dried prior to analysis. Adsorption and desorption data were collected in increments of 10% RH, ranging from 5% to 95% RH, under nitrogen purification. The equilibrium standard used for analysis exhibited a weight change of less than 0.0100% over 5 minutes, with a maximum equilibrium time of 3 hours. No initial moisture content correction data were available for the samples.

[0332] Thermogravimetric analysis (TGA)

[0333] Thermogravimetric analysis was performed using a Mettler-Toledo TGA / DSC3 analyzer. Temperature and enthalpy were regulated using indium, tin, and zinc, followed by verification with indium. Equilibrium was verified using calcium oxalate. Samples were placed in an open aluminum pot. The pot was then sealed, the lid punctured, and inserted into a TG furnace. A weighing aluminum pot configured as the sample pot was placed on a reference platform. The furnace was heated under nitrogen. Each sample was heated from ambient temperature to 350°C at a rate of 10°C / min. Although the thermal analysis plots are drawn at a reference temperature (x-axis), the results are reported based on the sample temperature.

[0334] X-ray powder diffraction (XRPD)

[0335] XRPD patterns were collected using a PANalytical X'Pert PRO MPD or PANalytical Empyrean diffractometer, which uses an incident beam of Cu radiation generated by a long, narrow focusing source. Cu Kα X-rays were focused through the sample and onto the detector using an elliptical gradient multilayer mirror. Prior to analysis, a silicon sample (NIST SRM 640e) was analyzed to verify that the observed Si 111 peak position was consistent with the NIST-certified position. The sample was sandwiched between thin films of 3 μm thickness and transmission geometry analysis was performed. A beam stopper, a short anti-scattering extension region, and an anti-scattering blade were used to minimize background from air. Soler slits for the incident and diffracted beams were used to minimize the broadening and asymmetry of axial divergence. Diffraction patterns were collected using a scanning position sensing detector (X'Celerator) at a distance of 240 mm from the sample and DataCollector software version 2.2b or 5.5.

[0336] Example 4: Salt Data

[0337] Crystalline and anhydrous SRR G-1 benzenesulfonate, camphorsulfonate, and naphthalenesulfonate were successfully isolated. All three were obtained as 1:1 stoichiometric salts. For these, the addition of seed crystals was crucial for providing high yields of colorless crystalline salts. The XRPD plots of the salts are shown in the figure. Figure 25 The free base type A in the sample was compared. The scale-up and characterization of the salt will be described in detail in the following sections.

[0338] SRR G-1 benzenesulfonate type A

[0339] SRR G-1 benzenesulfonate type A is an anhydrous 1:1 stoichiometric salt with a distinct melting initiation around 186°C. The disproportionation of this salt in water is not significant.

[0340] The single-crystal structure of SRR G-1 benzenesulfonate type A was successfully determined. Therefore, it will have a crystal size of approximately 0.23 × 0.09 × 0.04 mm. 3 Colorless needles of various sizes are mounted randomly on a polymer ring. This is used in an X-ray tube equipped with a copper anode microfocus seal. Preliminary examination and data collection were performed on a RigakuSuperNova diffractometer with a Decrispilatus3 R 200K hybrid pixel array detector. Using a set angle of 13177 reflections in the range of 4.2570° < θ < 77.0580°, the cell constant and orientation matrix for data collection were obtained by least-squares refinement. The space group was determined to be P21 by the program CrysAlisPro. Data was collected up to a maximum diffraction angle (2θ) of 155.242° at room temperature.

[0341] The frame was integrated with CrysAlisPro. A total of 26,894 reflections were collected, of which 10,520 were unique. Lorentz and polarization corrections were applied to the data. The linear absorption coefficient of Cu Kα radiation was 3.323 mm. -1 Empirical absorption correction was performed using CrysAlisPro. Transmission coefficients ranged from 0.837 to 1.000. Average equivalent reflection intensity. Based on intensity, the consistency factor for the average value was 3.3%.

[0342] The structure was resolved using a direct method with Shelxt. The remaining atoms were determined in a subsequent difference Fourier synthesis. The structure was refined using Shelxt-2014. Hydrogen atoms were refined independently. The structure was refined in full-matrix least squares by minimizing the following function:

[0343]

[0344] The weight w is defined as 1 / [δ] 2 (F o 2 )+(0.0401P) 2 +(0.3205P)], where P=(F o 2 +2F c 2 ) / 3. The scattering factor is taken from the "International Crystallographic Tables". Of the 10520 reflections used in the refinement, only those with an intensity greater than twice their uncertainty [I>2δ(I)], 9411, were used to calculate the fitting residual R. The final refinement loop includes 723 variable parameters, 1 constraint, and has converged to the following unweighted and weighted agreement factors:

[0345] 0.0348

[0346] 0.0874

[0347] The observed standard deviation per unit weight (goodness-of-fit) was 1.05. The electron density at the highest peak in the final difference Fourier transform was 0.311 e / Å. 3 The value of the smallest negative peak is -0.280 e / Å. 3 .

[0348] The crystal system is monoclinic, and the space group is P21. The cell parameters and calculated volume are: a = 14.1207(3) Å, b = 8.74139(11) Å, c = 21.5361(4) Å, α = 90°, β = 106.1889(19)°, γ = 90°, V = 2552.89(8) Å 3 The formula weight is 570.44 g mol. -1 Z=4, the calculated density is 1.484 g cm⁻¹ -3 Table 17 summarizes further details regarding the crystal data and crystallographic data collection parameters. The atomic shift ellipsoid diagram for benzenesulfonate type A is shown below. Figure 26 As shown, the asymmetric unit comprises two SRR G-1 cations and two benzenesulfonate anions. The -SO3 moiety is modeled using the rotational disorder of the two anions. Figure 27 XRPD plots of Cu radiation generation calculated using Mercury and atomic coordinates, space group, and cell parameters derived from single-crystal structures are provided and compared with experimental plots.

[0349] Table 17 Crystal data and data collection parameters for SRR G-1 benzenesulfonate type A

[0350]

[0351]

[0352] Table 18 lists the observed XRPD peaks of SRR G-1 benzenesulfonate type A.

[0353] Table 18

[0354]

[0355]

[0356]

[0357]

[0358] solution 1 The HNMR spectrum is consistent with that of SRR G-1 and the 1:1 stoichiometric salt of benzenesulfonic acid. Residual solvent is not obvious, consistent with the unsolvated form.

[0359] Figure 28Thermal analysis plots for SRR G-1 benzenesulfonate type A are provided. TGA confirms negligible weight loss up to 186 °C, consistent with the anhydrous form. DSC shows a sharp endothermic reaction starting near 186 °C. This event is likely due to the simultaneous occurrence of melting and decomposition. A small amount of endothermic reaction is also evident near 164 °C. The nature of this endothermic reaction is unknown.

[0360] The possibility of disproportionation in water was investigated. SRR G-1 benzenesulfonate type A was pulped in water for 4 days. Excess solids were recovered and reanalyzed by XRPD to obtain evidence of free base or benzenesulfonic acid. The recovered substance was SRR G-1 benzoate type A, indicating that disproportionation did not occur under the evaluated conditions.

[0361] The following describes the process for producing a 1-gram scale of SRR G-1 benzenesulfonate type A. 0.50 molar equivalents of benzenesulfonic acid monohydrate were added to a container containing 1.17 g of free SRR G-1 base type A. A small amount of SRR G-1 benzenesulfonate type A was added as a seed crystal. 7 mL of ethyl acetate was added, followed by sonication. The main portion of the solid dissolved, producing a yellow solution, but a white solid precipitated immediately thereafter. Another 3 mL of ethyl acetate was added to facilitate slurry transfer and filtration. The solid was recovered by vacuum filtration and washed with 4 mL of ethyl acetate, then incubated overnight under vacuum at room temperature. Approximately 0.99 g of SRR G-1 benzenesulfonate type A was obtained.

[0362] SRR G-1 Camphor Sulfonate Type A

[0363] SRR G-1 camphor sulfonate type A is an anhydrous 1:1 stoichiometric salt with an apparent melting point of 172°C.

[0364] The XRPD plot of SRR G-1 camphor sulfonate type A was successfully indexed, indicating that it is mainly composed of a single-crystal phase. Figure 29 The SRR G-1 camphor sulfonate type A has a triclinic unit cell that can accommodate two SRR G-1 cations and two camphor sulfonate anions. The calculated unit volume from the index is 737.9 Å. 3 Consistent with its anhydrous form, its calculated density is 1.451 g / cm³. -3 The XRPD spectrum also contains a small number of minor peaks unrelated to SRR G-1 camphor sulfonate type A, the known polymorph of free alkali, or (+)-(1S)-camphor-10-sulfonic acid. These additional peaks are... Figure 30 Highlighted with an asterisk.

[0365] Table 19 lists the observed XRPD peaks of SRR G-1 camphor sulfonate type A.

[0366] Table 19

[0367]

[0368]

[0369] The solution 1 The 1H NMR spectrum is consistent with the stoichiometric salts of SRR G-1 and (+)-(1S)-camphor-10-sulfonic acid at a 1:1 ratio. Residual solvent is not obvious, consistent with the unsolvated form.

[0370] Figure 31 The thermal analysis diagram of SRR G-1 camphor sulfonate type A is provided. TGA shows that the weight loss up to 171°C is negligible, consistent with the anhydrous form. DSC exhibits a sharp endothermic reaction, with an onset temperature close to 172°C. This event is likely due to the simultaneous occurrence of melting and decomposition.

[0371] The following describes the procedure for producing a 750 mg scale of SRR G-1 camphor sulfonate type A. 0.43 g of (+)-(1S)-camphor-10-sulfonic acid (less than 0.9 molar equivalents) was added to a container containing a suspension of 0.86 g of SRR G-1 free base type A and 10 mL of ethyl acetate, providing a yellow suspension containing a small amount of undissolved solids. Seed crystals of SRR G-1 camphor sulfonate type A were added, and the suspension was sonicated, immediately inducing precipitation. The sample was sonicated for an additional ~10 minutes and then placed in a slurry for approximately 1 hour. The white solids were recovered by vacuum filtration and washed with 2 mL of ethyl acetate, followed by overnight incubation under vacuum at room temperature. Approximately 0.75 g of SRR G-1 camphor sulfonate type A was obtained.

[0372] SRR G-1 Naphthalenesulfonate Type A

[0373] SRR G-1 naphthalenesulfonate type A is an anhydrous 1:1 stoichiometric salt with an apparent melting onset temperature of 194°C. Based on XRPD results of the aqueous slurry, disproportionation of this salt does indeed occur in water.

[0374] The XRPD plot of SRR G-1 naphthalene sulfonate type A was successfully indexed, indicating that it is mainly composed of a single-crystal phase. Figure 32 SRRG-1 naphthate type A has a monoclinic unit cell that can accommodate four SRRG-1 cations and four naphthalenesulfonate anions. The calculated unit volume based on the index is 707.3 Å. 3 Consistent with its anhydrous form, its calculated density is 1.457 g / cm³. -3The XRPD plot also contains a small, weak peak near 4.4° (2θ) that is unrelated to SRR G-1 naphthalene sulfonate type A, the known polymorph of the free base, or naphthalene-2-sulfonic acid.

[0375] Table 20 lists the observed XRPD peaks of SRR G-1 naphthalenesulfonate type A.

[0376] Table 20

[0377]

[0378]

[0379]

[0380] The solution 1 The 1H NMR spectrum is consistent with the stoichiometric salts of SRR G-1 and naphthalene-2-sulfonic acid. Residual solvent is not obvious, consistent with the unsolvated form.

[0381] Figure 33 Thermoanalytical plots of SRR G-1 naphthalenesulfonate type A are provided. TGA showed a weight loss of approximately 0.5% up to 193 °C. Most of the loss occurred above ~100 °C. As mentioned above, since no organic solvent was observed by NMR, this loss is considered to be due to the volatilization of approximately 0.2 mol / mol of water. This suggests that the salt may exhibit limited hygroscopicity. DSC showed a sharp endothermic reaction with an onset temperature close to 194 °C. This event was likely due to the simultaneous occurrence of melting and decomposition.

[0382] The possibility of disproportionation in water was investigated. SRR G-1 naphthalenesulfonate type A was pulped in water for 5 days. Excess solids were recovered and reanalyzed by XRPD to obtain evidence of free base or naphthalene-2-sulfonic acid. The recovered substance was free base type A, indicating disproportionation under the evaluated conditions.

[0383] The following describes the process for producing a 600 mg scale of SRR G-1 naphthalenesulfonate type A. Seed crystals of SRR G-1 naphthalenesulfonate type A and a molar equivalent of 0.39 g of naphthalene-2-sulfonic acid were added to a container containing a suspension of 0.73 g of SRR G-1 free base type A and 9 mL of ethyl acetate. The yellow suspension with a small amount of undissolved solids was sonicated, resulting in a white precipitate. The slurry was sonicated again for ~5 minutes, then the solids were recovered by vacuum filtration, washed with 2 mL of ethyl acetate, and vacuum dried overnight at room temperature. Approximately 0.63 g of SRR G-1 naphthalenesulfonate type A was obtained.

[0384] Example 5: YUMM1.7 proliferation assay

[0385] YUMM1.7 cells were thawed and cultured for at least one passage, then in DMEM containing 5% FBS (Invitrogen) and 1% antibiotic-antifungal agent (gibco) at 37°C and 5% CO2. Proliferation was assessed by seeding 15,000 cells into 12-well plates, with each test condition repeated five times. The medium and agents were replaced on day 2. On day 4, cells were trypsinized with 0.25 ml of 0.05% trypsin and EDTA (Invitrogen) for 5 minutes to separate them from the plates, mixed with 0.75 ml of medium, and counted using a hemocytometer.

[0386] Table 21 shows the mean cell counts after 4 days of growth in YUMM1.7 proliferation assays performed with each composition at 500 nM. The same data are shown graphically. Figure 34 The cell count was in the tens of thousands (i.e., 100 is approximately 1,000,000). The initial cell count was 15,000. The RSS G-1 doubling rate was roughly the same as that of the medium. Racemic G-1 decreased by about half the doubling rate observed in the medium. Surprisingly, the SRR G-1 decrease by at least 1 / 10 the doubling rate observed in the medium, rather than the expected only 1 / 4 the decrease caused by G-1.

[0387] Table 21

[0388]

[0389] Example 6: Preclinical Rat Pharmacokinetic Results

[0390] Plasma concentrations of SRR G-1 free base, SRR G-1 benzenesulfonate, and SRR G-1 naphthalenesulfonate in rats were determined after oral administration. Three fasted male rats were orally administered 10 mg / kg of SRR G-1 free base, SRR G-1 benzenesulfonate, or SRR G-1 naphthalenesulfonate in a suspension of 0.5% hydroxypropyl methylcellulose and 99.5% water. Plasma was separated at 0.5, 1, 2, 4, 8, and 24 hours after SRR G-1 administration, and plasma concentrations were determined by LC-MS / MS. The results are shown in Tables 22-24. The data are graphically represented as follows. Figures 35-37 As shown. Figure 38 The comparison of all three results is shown.

[0391] Table 22

[0392]

[0393] Table 23

[0394]

[0395] Table 24

[0396]

[0397] Example 7: ADME Toxicology of SRR G-1 and RSS G-1

[0398] ADME-Tox: In vitro absorption

[0399] Drug transporter (fluorescence inhibition)

[0400] The control percentage was calculated using the following equation. The inhibition percentage was obtained by subtracting the control percentage from 100. The IC50 value (the concentration at which the control value is maximally inhibited) was determined using the Hill equation via nonlinear regression analysis of the concentration-response curve.

[0401]

[0402] The compound reading is a single reading in the presence of the test compound. T1 is the average reading in the absence of the test compound. Background (for P-gp and BCRP) is the average reading in the presence of the reference inhibitor at the highest effective concentration. Background (for OATP1B1, OATP1B3, OAT1, OAT3, and OCT2) is the average reading in the absence of both the test compound and the substrate.

[0403] ADME-Tox: In vitro metabolism

[0404] Cytochrome P450 inhibition (detected by HPLC-UV / VIS and HPLC-MS / MS)

[0405] Peak areas corresponding to each substrate metabolite were recorded. The percentage of control activity was then calculated by comparing the peak areas obtained in the presence of the test compound with those obtained in the absence of the test compound. Subsequently, the percentage of inhibition was calculated by subtracting the percentage of control activity from 100 for each compound. The IC50 value (the concentration at which the half-maximal inhibition of the control value is achieved) was determined by nonlinear regression analysis of the concentration-response curves using Hill equation curve fitting.

[0406] Transporter inhibition results - When measured with 10 μM SRR G-1 or RSS G-1, the inhibition rates of the following transporters exceeded 50%. For SRR G-1: OATP1B1 82.5%. For RSS G-1: OCT2 53.2%, OATP1B1 91.2%, and OATP1B3 74.3%.

[0407] Cytochrome P450 inhibition results - When measured with 10 μM SRR G-1 or RSS G-1, the following substances showed inhibition rates exceeding 50%. For SRR G-1: CYP2D6 - 74.3% and CYP2C8 - 66.7%. For RSS G-1: CYP2C9 - 50.4%.

[0408] Cytochrome P450 induction results – Hepatocytes from three different human cell lines were incubated with SRR G-1 or RSSG-1 at 1 μM, 10 μM, and 100 μM. For SRR G-1: CYP1A2 was induced at 1 μM and 10 μM in only one of the three cell lines, while CYP3A4 was induced at 10 μM in two of the three cell lines. For RSS G-1: CYP1A2 was induced at 10 μM and 100 μM in two of the three cell lines.

[0409] Example 8: Off-target selectivity determination

[0410] GPCR cAMP regulation

[0411] Cell processing – cAMP Hunter cell lines were expanded from refrigerated storage according to standard procedures. Cells were seeded at a total volume of 20 μm into white-walled 384-well microplates and incubated at 37°C for the appropriate time prior to assay. Regulation of cAMP was determined using DiscoverXHitHunter cAMP XS+ assay.

[0412] Gs agonist form – To determine the agonist, cells were incubated with the sample to induce a reaction. The culture medium was aspirated from the cells and replaced with 15 μL of 2:1 HBSS / 10 mM Hepes:cAMP XS+Ab reagent. Intermediate dilutions of the sample stock were performed to generate a 4X sample in the assay buffer. 4.5 μL of the 4X sample was added to the cells and incubated at 37°C or room temperature for 30 or 60 minutes. The final assay medium concentration was 1%.

[0413] Gi agonist form - To determine the agonist, cells were placed in EC... 80 The reaction was induced by incubation with the sample in the presence of forskolin. The culture medium was aspirated from the cells and replaced with 15 μL of 2:1 HBSS / 10 mM Hepes:cAMP XS+Ab reagent. Intermediate dilutions of the sample stock were performed to obtain a solution containing 4X EC. 80 4X samples were prepared in Forskolin assay buffer. 4.5 μL of the 4X sample was added to the cells and incubated at 37°C or room temperature for 30 or 60 minutes. The final assay medium concentration was 1%.

[0414] Antagonist Forms - To determine the antagonist, cells were pre-incubated with the sample and then subjected to EC. 80 Agonist challenge was performed at a specific concentration. The culture medium was aspirated from the cells and replaced with 10 μL of 1:1 HBSS / Hepes:cAMP XS+Ab reagent. 5 μL of the 4X compound was added to the cells and incubated at 37°C or room temperature for 30 minutes. 4.5 μL of 4X EC was then added to the cells. 80 The agonist was added to the cells and incubated at 37°C or room temperature for 30 or 60 minutes. For Gi-coupled GPCRs, including EC... 80 Forskolin.

[0415] Signal detection – After incubation with the appropriate compound, a detection signal was generated by incubation with a 20 μL cAMP XS+ED / CL solution (cocktail) for 1 hour, followed by incubation with a 20 μL cAMP XS+EA reagent at room temperature for 3 hours. The microplate was read after the chemiluminescence signal was generated and detected using a PerkinElmer Envision™ instrument.

[0416] Data Analysis - The activity of the compounds was analyzed using the CBIS Data Analysis Suite (ChemInnovation, CA). For the Gs agonist mode assay, the percentage of activity was calculated using the following formula: %Activity = 100% x (mean RLU of test sample - mean RLU of medium control) / (mean RLU of MAX control - mean RLU of medium control). For the Gs antagonist mode assay, the percentage of inhibition was calculated using the following formula: %Inhibition = 100% x (1 - (mean RLU of test sample - mean RLU of medium control) / (EC... 80 The average RLU of the control group minus the average RLU of the mediator control group is used to calculate the percentage of activity. For Gi agonist mode assays, the percentage of activity is calculated using the following formula: %Activity = 100% x (1 - (average RLU of the test sample - average RLU of the MAX control group) / (average RLU of the mediator control group - average RLU of the MAX control group)). For Gi antagonist or negative allosteric mode assays, the percentage of inhibition is calculated using the following formula: %Inhibition = 100% x (average RLU of the test sample - EC) 80 (mean RLU of control) / (mean RLU of positive control of forscoring - EC) 80 The average RLU of the control group. For primary screens, the upper limit of the response percentage is 0% or 100%, where the calculated response percentage returns a negative value or a value greater than 100, respectively.

[0417] Calcium migration

[0418] Cell preparation – Expand cell lines from frozen stock according to standard procedures. Cells (10,000 cells / well) were seeded in 50 μL (200 cells / L) total volume into 384-well microplates with black walls and clear bottoms coated with poly-D-lysine and incubated at 37°C for the appropriate time prior to testing. DMSO concentration ≤0.2% for all readings.

[0419] Dye loading was performed in 1X dye loading buffer, which consisted of 1X dye (DiscoverX, Calcium No WashPLUS kit, catalog number 90-0091), 1X additive A, and 2.5 mM probenecid in HBSS / 20 mM Hepes. Probenecid was freshly prepared. Cells were dye-loaded prior to the test. The culture medium was aspirated from the cells and replaced with 25 μL of dye loading buffer. Cells were incubated at 37°C for 45 min, followed by incubation at room temperature for 20 min.

[0420] Agonist Formation – To determine the agonist, cells were incubated with the sample to induce a reaction. After dye loading, cells were removed from the incubator, and 25 μL of the 2X compound in HBSS / 20 mM Hepes was added using a FLIPR Tetra (MDS). The compound was added, and agonist activity was measured on the FLIPR Tetra. Calcium migration was monitored for 2 minutes, with baseline readings at 5 seconds.

[0421] Antagonist Form – Cells were pre-incubated with the sample, loaded with dye, transferred to FLIPR Tetra (MDS), and then challenged with an agonist at an EC80 concentration. Calcium migration was monitored for 2 minutes, with baseline readings at 5 seconds.

[0422] Data Analysis - FLIPR Readings - Calculate the area under the curve (AUC) over the entire two-minute reading period. Analyze the compound activity using the CBIS Data Analysis Suite (ChemInnovation, CA). For agonist mode assays, calculate the percentage of activity using the following formula: %Activity = 100% x (mean RFU of test sample - mean RFU of mediator control) / (mean MAX RFU of control ligand - mean RFU of mediator control). For antagonist mode assays, calculate the percentage of inhibition using the following formula: %Inhibition = 100% x (1 - (mean RFU of test sample - mean RFU of mediator control) / (EC...) 80 The average RFU of the control group (mean RFU of the median control)). For initial screening, the upper limit for the percentage of response is 0% or 100%, where the calculated percentage of response returns a negative value or a value greater than 100, respectively.

[0423] nuclear hormone receptors

[0424] Cell processing – The PathHunter NHR cell line was expanded from frozen stock according to standard procedures. Cells were seeded in 20 μL total volume into white-walled 384-well microplates and incubated at 37°C for an appropriate time prior to assay. The assay medium contained charcoal-dextran filtered serum to reduce the levels of present hormones.

[0425] Agonist Formation – To determine the agonist, cells were incubated with the sample to induce a reaction. Intermediate dilutions of the sample stock were performed to generate a 5X sample in the assay buffer. 3.5 μL of the 5X sample was added to the cells and incubated at 37°C or room temperature for 3–16 hours. The final assay medium concentration was 1%.

[0426] Antagonist Form - To determine the antagonist, cells were pre-incubated with the antagonist and then subjected to EC24. 80 Agonist challenge was performed at a concentration of 1%. Intermediate dilutions of the sample stock were performed to generate a 5X sample in assay buffer. 3.5 μL of the 5X sample was added to the cells and incubated at 37°C or room temperature for 60 minutes. The medium concentration was 1%. 4.5 μL of 6X EC in assay buffer was added... 80 The agonist was added to the cells and incubated at 37°C or room temperature for 3–16 hours.

[0427] Signal detection – A detection signal is generated by a single addition of 12.5 or 15 μL (50% v / v) of the PathHunter assay reagent mixture (cocktail), followed by incubation at room temperature for one hour. The microplate is read after the signal is generated and detected using a PerkinElmer Envision™ instrument.

[0428] Data Analysis - The activity of compounds was analyzed using the CBIS Data Analysis Suite (ChemInnovation, CA). For agonist mode assays, the percentage of activity was calculated using the following formula: %Activity = 100% x (mean RLU of test sample - mean RLU of mediator control) / (mean MAX control ligand - mean RLU of mediator control). For antagonist mode assays, the percentage of inhibition was calculated using the following formula: %Inhibition = 100% x (1 - (mean RLU of test sample - mean RLU of mediator control) / (EC) 80The average RLU of the control group minus the average RLU of the mediator control group. Note that for selection assays, the ligand response results in a decrease in receptor activity (inverse agonist with a constitutively active target). For those assays, inverse agonist activity is calculated using the following formula: % inverse agonist activity = 100% x ((average RLU of the mediator control group - average RLU of the test sample) / (average RLU of the mediator control group - average RLU of the MAX control group)). For initial screening, the upper limit for the percentage of response is 0% or 100%, where the calculated percentage of response returns a negative value or a value greater than 100, respectively.

[0429] KINOMEscan combined assay

[0430] Protein Expression – For most assays, kinase-tagged T7 phage strains were grown in parallel in 24-well plates in E. coli hosts derived from BL21 strain. E. coli were grown to the logarithmic phase and infected with T7 phage from frozen stock (multiple of infection = 0.4) and incubated at 32°C with shaking until lysed (90–150 min). The lysate was centrifuged (6,000 x g) and filtered (0.2 μm) to remove cell debris. The remaining kinases were produced in HEK-293 cells and subsequently labeled with DNA for qPCR detection.

[0431] Capture ligand generation – Streptavidin-coated magnetic beads were treated with biotinylated small molecule ligands at room temperature for 30 minutes to generate affinity resins for kinase assays. The ligand beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligands and reduce nonspecific phage binding.

[0432] Binding reaction assembly – The binding reaction was assembled by binding the kinase, ligand affinity beads, and test compound in 1X binding buffer (20% SeaBlock, 0.17X PBS, 0.05% Tween 20, 6 mM DTT). All reactions were performed in 0.02 mL final volumes of polypropylene 384-well plates. The plates were incubated at room temperature with shaking for 1 hour, and the affinity beads were washed with wash buffer (1x PBS, 0.05% Tween 20). The beads were then resuspended in elution buffer (1x PBS, 0.05% Tween 20, 0.5 μM non-biotinylated affinity ligand) and incubated at room temperature with shaking for 30 minutes. The kinase concentration in the elution buffer was measured by qPCR.

[0433] Signal detection – kinase concentration in the elution buffer was measured by qPCR. The qPCR reaction was assembled by adding 2.5 μL of kinase elution buffer to 7.5 μL of a qPCR master mix containing 0.15 μM amplicon primers and 0.15 μM amplicon probes. The qPCR protocol consisted of 35 cycles: a 10-minute hot start at 95°C, followed by 15 seconds at 95°C and 1 minute at 60°C.

[0434] Data Analysis - Calculating Percentage of Reaction

[0435]

[0436] Test compound = SRR G-1

[0437] Negative control = DMSO (100% control)

[0438] Positive control = control compound (0% control)

[0439] Use the following formula to convert the control percentage to the response percentage: Response percentage = (100 - Control percentage). For initial screening, the response percentage is capped at 0% or 100%, where the calculated response percentage returns a negative value or a value greater than 100, respectively.

[0440] Data Analysis - Combining Constants (Kds)

[0441] Using the Hill equation with the constant (Kds) as a reference, the response curve was calculated from the standard dose curve:

[0442]

[0443] The Hill slope is set to -1.

[0444] The curves were fitted using nonlinear least squares fitting and the Levenberg-Marquardt algorithm.

[0445] Ion channel assay

[0446] Cell processing – Expand cell lines from frozen stock according to standard procedures. Seed cells in 20 μL total volume into 384-well microplates with black walls and clear bottoms coated with poly-D-lysine and incubated at 37°C for the appropriate time prior to testing.

[0447] Dye loading – Where applicable, the assay is performed in a 1X dye loading buffer consisting of 1X dye and 2.5 mM probenecid. Probenecid is freshly prepared. Load cells with the dye prior to the assay. Incubate cells at 37°C for 30–60 minutes.

[0448] Agonist / Initiator (Opener) Form - To determine the agonist, cells are incubated with the sample to induce a reaction. Intermediate dilutions of the sample stock are performed to generate 2-5X samples in the assay buffer. 10-25 μL of the 2-5X sample is added to the cells and incubated at 37°C or room temperature for 30 minutes. The final assay medium concentration is 1%.

[0449] Antagonist / Blocker Formation - To determine the antagonist, cells are pre-incubated with the sample. Intermediate dilutions of the sample stock are performed to generate 2-5X samples in assay buffer. After dye loading, cells are removed from the incubator and, where appropriate, placed on EC. 80 Add 10–25 μL of 2–5X sample to the cells in the presence of the agonist. Incubate the cells in the dark at room temperature for 30 minutes to equilibrate the plate temperature. The medium concentration is 1%.

[0450] Signal Detection - Measuring Compound Activity on FLIPR Tetra (MDS)

[0451] Data Analysis - The activity of compounds was analyzed using the CBIS Data Analysis Suite (ChemInnovation, CA). For agonist mode assays, the percentage of activity was calculated using the following formula: %Activity = 100% x (mean RLU of test sample - mean RLU of mediator control) / (mean MAX control ligand - mean RLU of mediator control). For antagonist inhibition percentages, the following formula was used: %Inhibition = 100% x (1 - (mean RLU of test sample - mean RLU of mediator control) / (EC) 80 The average RLU of the control group is equal to the average RLU of the medium control group. For the initial screening, the upper limit of the response percentage is 0% or 100%, where the calculated response percentage returns a negative value or a value greater than 100, respectively.

[0452] Transporter determination

[0453] Cell processing – Expand cell lines from frozen stock according to standard procedures. Seed cells in 25 μL total volume into 384-well microplates with black walls and clear bottoms coated with poly-D-lysine and incubated at 37°C for the appropriate time prior to testing.

[0454] In the form of the inhibitor / antagonist, after cell plating and incubation, the culture medium was removed and 25 μL of the 1X compound in 1X HBSS / 0.1% BAS was added. The compound was then incubated with the cells at 37°C for 30 minutes.

[0455] Dye loading – The assay was performed in a 1X dye loading buffer consisting of 1X dye, 1X HBSS / 20 mM Hepes. After compound incubation, 25 μL of 1X dye was added to the wells. Cells were incubated at 37°C for 30–60 minutes.

[0456] Signal detection - After dye incubation, the microplate is transferred to a PerkinElmer Envision™ instrument for fluorescence signal detection.

[0457] Data Analysis – The activity of the compounds was analyzed using the CBIS Data Analysis Suite (ChemInnovation, CA). For the antagonist mode assay, the percentage of inhibition was calculated using the following formula: %Inhibition = 100% x (1 - (mean RLU of test sample - mean RLU of medium control) / (mean RLU of positive control - mean RLU of medium control)). For preliminary screening, the upper limit for the percentage of response was 0% or 100%, where the calculated percentage of response returned a negative value or a value greater than 100, respectively.

[0458] Enzyme assay

[0459] Enzyme preparations - Enzyme preparations are sourced from various suppliers - AChE (R&D Systems), COX1 and COX2 (BPSBioscience), MAOA (Sigma), PDE3A and PDE4D2 (Signal Chem).

[0460] Enzyme Activity Assays – Enzyme activities are determined by measuring the amount of substrate consumed or the amount of product produced over time. Different detection methods are used to measure the concentrations of substrate and product in each enzymatic assay. AChE: Before adding the substrate, the enzyme and test compound are pre-incubated at room temperature for 15 minutes. Thioacetylcholine and DTNB are added and incubated at room temperature for 30 minutes. The signal is detected by measuring the absorbance at 405 nm. COX1 and COX2: The enzyme stock is diluted in assay buffer (40 mM Tris-HCl, 1X PBS, 0.5 mM phenol, 0.01% Tween-20 + 100 nM hemoglobin) and equilibrated with the compound at room temperature for 30 minutes (binding incubation). Arachidonic acid (1.7 μM) and Ampliflu Red (2.5 μM) are prepared and dispensed into reaction plates. The plates are immediately read on a fluorometer, with emission detection at 590 nm and excitation wavelength at 544 nm. MAOA: Before adding the substrate, pre-incubate the enzyme and test compound at 37°C for 15 minutes. Initiate the reaction by adding kynuramine and incubating at 37°C for 30 minutes. Terminate the reaction by adding NaOH. Determine the amount of 4-hydroquinoline formed using fluorescence spectroscopy with emission detection at 380 nm and excitation at 310 nm. PDE3A and PDE4D2: Before adding the substrate, pre-incubate the enzyme and test compound at room temperature for 15 minutes. Add cAMP substrate (concentration equal to EC50). 80 Incubate at room temperature for 30 minutes. Terminate the enzyme reaction by adding 9 mM IBMX. Detect the signal using the HitHunter® cAMP assay kit.

[0461] Signal detection - For each measurement, transfer the microplate to the PerkinElmer Envision™ instrument and take the reading as described.

[0462] Data Analysis – The activity of compounds was analyzed using the CBIS Data Analysis Suite (ChemInnovation, CA). For enzyme activity assays, the percentage of inhibition was calculated using the following formula: %Inhibition = 100% x (1 - (mean RLU of test sample - mean RLU of medium control) / (mean RLU of positive control - mean RLU of medium control)). For preliminary screening, the upper limit for the percentage of reaction was 0% or 100%, where the calculated percentage of reaction returned a negative value or a value greater than 100, respectively.

[0463] Results of off-target selectivity analysis

[0464] The selectivity of SRR G-1 and RSS G-1 against potential off-target effects was tested in 78 assays at concentrations up to 10 μM in dose-response form. SRR G-1 exhibited selectivity only against the following measurable IC50 values. 50or EC 50 The concentrations were 2.5 μM for cannabinoid receptor 1, 8.2 μM for HTR2B, 0.87 μM for OPRD1, and 6.68 μM for OPRM1. RSS G-1 only exhibited measurable IC50 values ​​for the following: 50 or EC 50 The concentrations were 3.1 μM for cannabinoid receptor 1, 2.07 μM for ADRA2A, 2.1 μM for HTR1A, and 4.76 μM for AR.

Claims

1. A compound comprising the following formula: 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl) ethyl-1-one or a derivative thereof, wherein the chiral purity of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl) ethyl-1-one or a derivative thereof is about 90% or higher.

2. The compound of claim 1, wherein the compound is a crystal as demonstrated by XRPD analysis or an amorphous substance or a mixture of crystals and amorphous substances as demonstrated by XRPD analysis.

3. The compound of claim 1, wherein the chiral purity of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadieno[c]quinolin-8-yl) ethyl-1-one or a derivative thereof is substantially free of its opposite enantiomers.

4. The compound of claim 1, wherein the form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)acetoone is selected from crystal form A, characterized in that... XRPD plots having peaks at approximately 2θ (±0.20) at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86; Crystal form B, characterized by XRPD plots having peaks at approximately 2θ (±0.20) at approximately 13.98, approximately 15.44, approximately 19.67, approximately 21.55, and approximately 22.05; Crystal form C, characterized by XRPD plots having peaks at approximately 2θ (±0.20) at approximately 10.73, approximately 12.77, approximately 13.49, approximately 16.09, and approximately 20.60; amorphous; or combinations thereof.

5. The compound of claim 1, wherein the crystal form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)acet-1-one is selected from crystal form A, characterized in that... XRPD plots having peaks at approximately 2θ (±0.20) at approximately 5.75, approximately 20.54, approximately 20.71, approximately 21.25, and approximately 21.86; crystal form B, characterized in that XRPD plots have peaks at approximately 13.98, approximately 15.44, approximately 19.67, approximately 21.55, and approximately 22.05 at approximately 2θ (±0.20); crystal form C, characterized in that XRPD plots have peaks at approximately 10.73, approximately 12.77, approximately 13.49, approximately 16.09, and approximately 20.60 at approximately 2θ (±0.20); or combinations thereof.

6. The compound of claim 5, wherein the crystal form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)acet-1-one is crystal form A, characterized in that... The XRPD plot has peaks located at approximately 5.75, 20.54, 20.71, 21.25, and 21.86, expressed in degrees 2θ (±0.20).

7. The compound of claim 6, wherein the crystalline form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)ethyl-1-one is crystalline form A, which is further characterized by an XRPD plot having peaks at degrees 2θ (±0.20) at the following values: about 5.75, about 9.56, about 10.53, about 17.03, about 20.54, about 20.71, about 21.25, about 21.86, about 24.67, and about 28.

06.

8. The compound of claim 7, wherein the crystalline form of 1-((3aS,4R,9bR)-4-(6-bromobenzo[d][1,3]dioxacyclopenten-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopentadien[c]quinolin-8-yl)ethyl-1-one is crystalline form A, further characterized by an XRPD plot having peaks in degrees 2θ (±0.20) at the following locations: about 5.75, about 9.56, about 10.53, about 10.81, about 13.02, about 14.66, about 14.79, about 16.23, about 17.03, about 20.54, about 20.71, about 21.25, about 21.86, about 24.67, and about 28.

06.

9. The compound of claim 1, wherein the derivative thereof is a salt or a eutectic.

10. The compound of claim 1, wherein the derivative thereof is selected from salts or cocrystals formed with benzenesulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, ethane-1,2-disulfonic acid, hydrochloric acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, sulfuric acid, p-toluenesulfonic acid, or combinations thereof.

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