Methods for treating cancers associated with ETBR activation

Abstract

Disclosed herein are methods for treating cancer with a combination therapy comprising a CAR-expressing immune cell (e.g., CAR-T cell) and an ETBR antagonist. Also disclosed herein are methods for treating cancer with a combination therapy comprising a CAR-expressing immune cell (e.g., a CAR-T cell), an immune checkpoint inhibitor (e.g., an anti-PD1 antibody), and an ETBR antagonist.

Description

[0001] Overview

[0002] In some embodiments, this document discloses a method for treating a subject with a target cancer in need, the method comprising administering to the subject a combination comprising: (a) immune cells expressing a chimeric antigen receptor (CAR), wherein the CAR specifically binds to the target; and (B) an effective amount of an ETBR antagonist.

[0003] In various embodiments of a method for treating cancers expressing a target in subjects with appropriate need using immune cells expressing a chimeric antigen receptor (CAR) (wherein the CAR specifically binds to the target), an improvement is disclosed herein that includes co-administering an effective amount of an ETBR antagonist to the subject.

[0004] In some embodiments, this document discloses a method for treating a subject with a target cancer in need, the method comprising administering to the subject a combination comprising: (a) immune cells expressing a chimeric antigen receptor (CAR), wherein the CAR specifically binds to the target; (b) an immune checkpoint inhibitor (ICPI); and (c) an effective amount of an ETBR antagonist.

[0005] In various embodiments of a method for treating cancers expressing a target in subjects with appropriate need using immune cells expressing a chimeric antigen receptor (CAR) (wherein the CAR specifically binds to the target) and immune checkpoint inhibitors (ICPIs), an improvement is disclosed herein that includes co-administering an effective amount of an ETBR antagonist to the subject.

[0006] In some implementations, the target is a tumor-associated antigen. In some implementations, the tumor-associated antigen is HER2.

[0007] In some embodiments, this document discloses a method for treating HER2-expressing cancer in a subject with a corresponding need, the method comprising administering to the subject a combination comprising: (a) immune cells expressing a chimeric antigen receptor (CAR), wherein the CAR specifically binds to HER2, and (b) an effective amount of an ETBR antagonist.

[0008] In various embodiments of a method for treating HER2-expressing cancers in subjects with appropriate need using immune cells expressing chimeric antigen receptors (CARs) (wherein the CARs specifically bind to HER2), an improvement is disclosed herein that includes co-administering an effective amount of an ETBR antagonist to the subject.

[0009] In some embodiments, the ICPI is an anti-PD1 antibody, an anti-PD-L1 antibody, an anti-CTLA4 antibody, or a variant or functional fragment thereof. In some embodiments, the ICPI is an anti-PD1 antibody.

[0010] In some implementations, the ETBR antagonist is a deuterated ETBR antagonist.

[0011] In some implementations, the deuterated ETBR antagonist is a compound of formula (1):

[0012]

[0013] Or its stereoisomers, wherein:

[0014] n is an integer between 0 and 4;

[0015] m is an integer between 0 and 3;

[0016] X is a positively charged equilibrium ion;

[0017] R1 and R3 are independently -H, -D, -CH3, -CH2D, -CHD2, or -CD3;

[0018] R 2a R 2b R4, R5, and R6 are independently -CH3, -CH2D, -CHD2, or -CD3; and

[0019] R1, R 2a R 2b Or at least one of R3 contains deuterium.

[0020] In some embodiments, the compound is a compound of formula (1a):

[0021]

[0022] Or its stereoisomers.

[0023] In some implementations, the compound is a compound of formula (2):

[0024]

[0025] Or its stereoisomers.

[0026] In some embodiments, the compound is a compound of formula (2a):

[0027]

[0028] Or its stereoisomers.

[0029] In some implementations, the compound is a compound of formula (3):

[0030]

[0032] Or its stereoisomers.

[0033] In some embodiments, the compound is a compound of formula (3a):

[0034]

[0035] Or its stereoisomers.

[0036] In some embodiments, the compound is a compound of formula (4):

[0037]

[0038] Or its stereoisomers.

[0039] In some embodiments, the compound is a compound of formula (4a):

[0040]

[0041] Or its stereoisomers.

[0042] In some implementations, the compound is a compound of formula (5):

[0043]

[0044] Or its stereoisomers.

[0045] In some embodiments, the compound is a compound of formula (5a):

[0046]

[0047] Or its stereoisomers.

[0048] In some implementations, the compound is a compound of formula (6):

[0049]

[0050] Or its stereoisomers.

[0051] In some embodiments, the compound is a compound of formula (6a):

[0052]

[0054] Or its stereoisomers.

[0055] In some implementations, n is 1, R1 is -D, and R2a and R 2b Each is -CH3.

[0056] In some implementation schemes, R 2a It is -CH3, and R 2b It is -CH2D.

[0057] In some implementation schemes, R 2a and R 2b Each is -CH2D.

[0058] In some implementations, n is 1, R1 is -D; and R 2a and R 2b Each is -CH2D.

[0059] In some implementations, the deuterated ETBR antagonist is:

[0060] BQ-788-B

[0061]

[0062] BQ-788-A

[0063]

[0064] BQ-788-C

[0065]

[0066] Or its stereoisomers.

[0067] In some implementations, the ETBR antagonist is:

[0068] BQ-788-B

[0069]

[0070] Or its stereoisomers.

[0071] In some implementations, the ETBR antagonist is in a pharmaceutically acceptable excipient containing dimethyl sulfoxide (DMSO).

[0072] In some implementations, the immune cells expressing CAR are CAR-T cells or CAR-NK cells. In some implementations, the immune cells expressing CAR are CAR-T cells. In some implementations, the cancer is selected from the group consisting of: breast cancer, gastric cancer, esophageal cancer, ovarian cancer, endometrial cancer, bladder cancer, lung cancer, cervical cancer, colon cancer, and head and neck cancer.

[0073] In some implementations, the cancer is ETBR-related cancer.

[0074] In some implementation schemes, ETBR-related cancers are selected from the group consisting of: breast cancer, stomach cancer, esophageal cancer, ovarian cancer, endometrial cancer, bladder cancer, lung cancer, cervical cancer, colon cancer, and head and neck cancer.

[0075] In some implementations, the cancer is cancer that expresses HER2.

[0076] In some implementations, cancers that express HER2 are HER2+ cancers.

[0077] In some implementations, cancers that express HER2 are low-HER2 cancers.

[0078] In some implementations, cancers expressing HER2 are selected from the group consisting of: breast cancer, stomach cancer, esophageal cancer, ovarian cancer, endometrial cancer, bladder cancer, lung cancer, cervical cancer, colon cancer, and head and neck cancer. Brief description of the attached diagram

[0080] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several embodiments of the invention and, together with the specification, serve to explain the principles of the invention. The drawings are for illustrative purposes only and should not be construed as limiting the invention. Other objects, features, and advantages of the invention will become apparent from the following detailed description taken in conjunction with the accompanying drawings, which illustrate illustrative embodiments of the invention, in which:

[0081] Figure 1 The endothelin B receptor (ETBR) cellular signaling pathway is illustrated. ETBR is a seven-transmembrane G protein-coupled receptor (GPCR). Endothelin-1 (ET-1) is the ligand of ETBR. Binding of ET-1 to the receptor leads to the activation of many downstream kinases, including PTK, RAF, MEK, and MAPK / ERK.

[0082] Figure 2 The tumor volume over time in 4T1 transplanted mice treated with indicated combination therapy was depicted.

[0083] Figure 3A and Figure 3B Tumor growth of 4T1 tumors was depicted from day 11 to day 17 post-transplantation in mice treated with indicated combination therapy. Figure 3A The tumor volume over time was depicted. Figure 3B Tumor volume was depicted on day 17 post-transplantation. The differences between group 8 and groups 1 and 2 were statistically significant (P < 0.01). The differences between group 8 and groups 3 and 4 were statistically significant (P < 0.05).

[0084] Figure 4Tumor volume was depicted on day 29 post-transplantation of 4T1 tumors in mice treated with the indicated combination therapy. Differences between groups 1 and 2 were P<0.05. Differences between groups 1 and 4 were P<0.01. Differences between groups 4 and 6 were P<0.01. Differences between groups 1 and 5, 6, 7, and 8 were P<0.001. Differences between groups 2 and 5, 6, 7, and 8 were P<0.001. Differences between groups 3 and 5, 6, 7, and 8 were P<0.001. Differences between groups 4 and 5, 7, and 8 were P<0.001.

[0085] Figure 5A and Figure 5B Tumor growth at the experimental endpoint (day 30 post-transplantation of 4T1 tumor) was depicted in mice treated with the indicated combination therapy. Figure 5A An image of the harvested tumor is shown. Figure 5B Tumor weight was described on day 30 post-transplantation. The difference between group 1 and group 4 was P<0.05. The differences between group 4 and groups 5, 6, and 7 were P<0.05, respectively. The difference between group 4 and group 8 was P<0.01. The differences between group 3 and groups 6 and 7 were P<0.01. The differences between group 1 and groups 5, 6, 7, and 8 were P<0.001, respectively. The differences between group 2 and groups 5, 6, 7, and 8 were P<0.001, respectively. The differences between group 3 and groups 5 and 8 were P<0.001, respectively.

[0086] Detailed Explanation

[0087] This document discloses combination therapies and methods for treating cancer with a single dosage form, administered simultaneously or separately, comprising an ETBR antagonist as described herein and CAR-expressing immune cells (e.g., CAR-T cells). In some embodiments, the treatment combination also comprises an immune checkpoint inhibitor (ICPI), such as an anti-PD1 antibody or an anti-PD-L1 antibody. In some embodiments, the ETBR antagonist is a specific deuterated ETBR antagonist as described herein. In some embodiments, the specific deuterated ETBR antagonist is deuterated BQ-788 (BQ-788-A, BQ-788-B, or BQ-788-C, or combinations thereof).

[0088] In some embodiments, the combination therapy described herein exhibits a synergistic effect because the pharmaceutical composition achieves at least one of the following: a greater therapeutic effect (i.e., more effective) than the additive therapeutic effect obtained by administering the individual components, a greater therapeutic effect than the therapeutic effect achieved by administering a higher dose of the individual components, a similar or greater therapeutic effect observed relative to administering the individual components but with fewer adverse events or side effects (i.e., an improved therapeutic window), or an increased duration of effect, or a similar or greater therapeutic effect at a lower dose of one or both components, or a combination thereof.

[0089] definition

[0090] 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 to which this invention pertains. The terminology used in this specification is for describing particular embodiments only and is not intended to limit the invention.

[0091] When a range of values ​​is provided, it should be understood that unless the context explicitly specifies otherwise (e.g., in the case of a group containing multiple carbon atoms, the number of each carbon atom falling within the range is provided), every intermediate value between the upper and lower limits of the range up to one-tenth of the lower limit unit, as well as any other specified value or intermediate value within the range, is included within the scope of this invention. The upper and lower limits of these smaller ranges may be independently included within the smaller ranges, and also within the scope of this invention, subject to any specifically excluded limits within the ranges. Where the range includes one or both limit values, the range excluding one of these included limit values ​​is also included in this disclosure.

[0092] Unless the context clearly indicates otherwise, the articles “a” and “an” as used herein and in the appended claims are used to refer to one or more (i.e., at least one) articles. As an example, “an element” means one or more elements.

[0093] The phrase “and / or” as used herein in the specification and claims should be understood to mean “one or two” of the elements so connected (i.e., elements that are jointly present in some cases and separately present in others). The use of “and / or” to list multiple elements should be interpreted in the same way, i.e., “one or more” of the elements so connected. In addition to the elements specifically indicated by the “and / or” clause, other elements may optionally be present, whether related to or unrelated to those specifically indicated. Thus, as a non-limiting example, references to “A and / or B” used in conjunction with open-ended language (such as “comprising / including”) may, in one embodiment, refer to only A (optionally including elements other than B); in another embodiment, refer to only B (optionally including elements other than A); in yet another embodiment, refer to both A and B (optionally including other elements); and so on.

[0094] As used herein in the specification and claims, “or” should be understood to have the same meaning as “and / or” as defined above. For example, when separating items in a list, “or” or “and / or” should be interpreted as inclusive, i.e., including multiple elements or at least one of the elements in the list, but also including more than one, and optionally including additional unlisted items. Terms that explicitly indicate the opposite, such as “only one” or “exactly one”, or “consisting of” as used in the claims, will refer to including multiple elements or exactly one element in the list of elements. In general, when preceded by exclusive terms such as “any,” “one of,” “only one of,” or “exact one of,” the term “or” as used herein should only be interpreted as indicating an exclusive alternative (i.e., “one or the other, but not both”).

[0095] In the claims and the foregoing description, all transitional phrases such as “comprising / including,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” “comprising,” etc., should be understood as open-ended, meaning including but not limited to. As described in Section 2111.03 of the U.S. Patent Examination Procedure Manual, only the transitional phrases “consisting of…” and “consisting substantially of…” should be closed or semi-closed transitional phrases, respectively.

[0096] As used herein and in the claims, the phrase “at least one” referring to a list of one or more elements should be understood to mean at least one element selected from any one or more elements in the list of elements, but not necessarily including each and at least one of every elements specifically listed in the list of elements, and does not exclude any combination of elements in the list of elements. This definition also allows for the optional presence of elements other than those specifically indicated in the list of elements referred to by the phrase “at least one,” whether related to or unrelated to those specifically indicated elements. Thus, as a non-limiting example, “at least one of A and B” (or equivalently, “at least one of A or B”, or equivalently, “at least one of A and / or B”) in one embodiment may refer to at least one A, optionally including more than one A, while B is absent (and optionally including elements other than B); in another embodiment, it may refer to at least one B, optionally including more than one B, while A is absent (and optionally including elements other than A); in yet another embodiment, it may refer to at least one A, optionally including more than one A, and at least one B, optionally including more than one B (and optionally including other elements); and so on.

[0097] It should also be understood that, unless the context otherwise requires, in some methods described herein that include more than one step or action, the order of the steps or actions of the method is not limited to the order in which the steps or actions of the method are listed.

[0098] The term "combination therapy" refers to both simultaneous administration (administration of two or more therapeutic agents at the same time) and simultaneous administration (administration of one or more therapeutic agents at a time different from the administration of another one or more therapeutic agents). In some embodiments, the therapeutic agents are present in the patient simultaneously to a certain extent, for example, in an effective amount. In some embodiments, one or more compounds described herein are administered in combination with other bioactive agents, particularly including anticancer agents. In some embodiments, the combination therapy of compounds produces synergistic activity, including anticancer activity.

[0099] In this context, unless otherwise stated, the term "compound" as used herein means any specific compound disclosed herein and includes tautomers, positional isomers, geometric isomers, stereoisomers where applicable (including their optical isomers (enantiomers) and other stereoisomers (diastereomers)), and pharmaceutically acceptable salts and derivatives (including prodrug forms) where applicable. In the context of its use, the term "compound" generally refers to a single compound, but may also include other compounds, such as stereoisomers, positional isomers, and / or optical isomers (including racemic mixtures), and specific enantiomers of the disclosed compound or mixtures enriched with enantiomers. In this context, the term also refers to a prodrug form of the compound that has been modified to facilitate the administration and delivery of the compound to the active site. It should be noted that many substituents and variables, etc., associated with the compounds of the present invention are described. Those skilled in the art will understand that the molecules described herein are stable compounds as generally described below. When bonds are shown, both double and single bonds are represented in the context of the compounds shown.

[0100] As used herein, the terms “treat,” “treating,” and “treatment” refer to any action that provides benefit to a patient to whom the compounds of the present invention may be administered, including treatment of any disease state or condition regulated by a protein to which the compounds of the present invention bind. Disease states or conditions, including cancer, that can be treated with the compounds according to this disclosure are described above.

[0101] The term "pharmaceutically acceptable salt" is used throughout the specification to describe, where applicable, the salt form of one or more compounds described herein, which enhances the solubility of the compound in the gastric juices of a patient's gastrointestinal tract, thereby promoting the solubility and bioavailability of the compound. Pharmaceutically acceptable salts include, where applicable, those derived from pharmaceutically acceptable inorganic or organic bases and acids. Suitable salts include salts derived from alkali metals (such as potassium and sodium), alkaline earth metals (such as calcium and magnesium), and ammonium salts. In some embodiments, sodium and potassium salts are suitable neutralizing salts of phosphates.

[0102] The term "pharmaceuticalally acceptable derivative" is used throughout the specification to describe any pharmaceutically acceptable prodrug form (such as ester, amide, or other prodrug group) that directly or indirectly provides a compound of the invention or an active metabolite of the compound of the invention when administered to a patient.

[0103] The term "effective" is used to describe the amount of a compound, composition, or component that, when used in the context of its intended use, achieves the desired result. The term "effective" includes all other effective amount or effective concentration terms otherwise described or used in this application.

[0104] The term “therapeutic effective amount” refers to the amount that is sufficient to achieve the treatment as defined herein when applied to a mammal that requires such treatment.

[0105] The terms “patient” or “subject” are used throughout the specification to describe an animal (e.g., a human or domestic animal) to which it is provided with treatment (including prophylactic treatment) using the compositions according to this disclosure. For treatments of infections, conditions, or disease states specific to a particular animal (such as a human patient), the term “patient” refers to that particular animal, including domestic animals (such as dogs or cats) or farm animals (such as horses, cattle, sheep, etc.). Generally, in this disclosure, the term “patient” refers to a human patient unless otherwise stated or implied in the context of its use. Activation of the ETBR by endothelins such as ET-1 and ET-3 leads to a variety of molecular events that promote the invasion and metastasis of melanoma. Without being bound by any particular theory, it is presumed that although most melanomas express ETBR, a subset of these melanomas also express ETBR activators ET-1 and / or ET-3. Therefore, this subset is most likely to depend on ETBR activation for survival, invasive potential, and metastatic potential. Therefore, patients in this subset are most likely to respond to ETBR blockade. Furthermore, this subset of patients is the least likely to respond to immune-based therapies.

[0106] Deuterated ETBR antagonists

[0107] In some embodiments, the methods and pharmaceutical compositions described herein use or comprise a deuterated ETBR antagonist, such as the deuterated form of BQ-788 as described herein. The specific deuterated compounds mentioned herein are advantageous because they offer one or more improvements in pharmaceutical properties (e.g., efficacy, solubility) compared to the non-deuterated form of the same compound. The specific deuterated ETBR antagonists mentioned herein, including the deuterated forms of BQ-788 (e.g., BQ-788A, BQ-788-B, and BQ-788-C), are described in WO2019140324, which is incorporated herein by reference in its entirety. The specific deuterated ETBR antagonist BQ-788-B is also known as ENB-003.

[0108] Endothelin B receptor (ETBR) pathway Figure 1Endothelin-1 plays a crucial role in the metastatic spread of melanoma and is therefore a target for therapeutic intervention. The endothelin-B receptor is a 7-transmembrane G protein-coupled receptor (GPCR). It is expressed at very low levels in normal melanocytes but is upregulated during melanoma development and progression. Current melanoma drug targets, RAF and MEK kinases, are activated by deuterated ETBR. In some implementations, the physiological role of ETBR is to clear excess levels of endothelin-1 (ET-1) from circulation. Without being bound by any particular theory, it is hypothesized that administration of an ETBR antagonist (e.g., a specific deuterated ETBR antagonist) prevents ET-1 clearance and increases serum ET-1 levels. Elevated serum ET-1 levels are associated with a variety of adverse reactions, including hypertension, pulmonary hypertension, and renal vasoconstriction, due to its activation of the endothelin A receptor (ETAR).

[0109] In some embodiments, this specification provides a composition comprising an ETBR antagonist (e.g., a specific deuterated ETBR antagonist, such as the deuterated form of BQ-788 as described herein) and a pharmaceutically acceptable carrier. In some embodiments, this specification provides a composition, such as a pharmaceutical composition, comprising an effective amount of an ETBR antagonist (e.g., a specific deuterated ETBR antagonist, such as the deuterated form of BQ-788 as described herein) and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition described herein may be a unit dosage form configured for single or multiple administrations, such as once or multiple times daily, weekly, or monthly.

[0110] In some implementations, the specific deuterated ETBR antagonist is a compound of formula (1):

[0111]

[0112] in

[0113] n is an integer between 0 and 5;

[0114] m is an integer between 0 and 3;

[0115] X is a positively charged equilibrium ion;

[0116] R1 and R3 are independently -H, -D, -CH3, -CH2D, -CHD2, or -CD3;

[0117] R 2a R 2b R4, R5, and R6 are independently -CH3, -CH2D, -CHD2, or -CD3; and

[0118] At least one of R1, R2, or R3 contains deuterium.

[0119] In some embodiments, the specific deuterated ETBR antagonist of formula (1) contains 1-8 deuterium atoms. In specific embodiments, the specific deuterated ETBR antagonist of formula (1) contains 1, 2 or 3 deuterium atoms.

[0120] In some embodiments, the compound has the following formula (1a):

[0121]

[0122] Where X and R 2a R 2b R4, R5, and R6 are as described in this article.

[0123] In some implementations, the specific deuterated ETBR antagonist is a compound of formula (2):

[0124]

[0125] Where X, m, n, R1, R3, R 2a R 2b R4, R5, and R6 are as described in this article.

[0126] In some embodiments, the compound has the following formula (2a):

[0127]

[0128] Where X and R 2a R 2b R4, R5, and R6 are as described in this article.

[0129] In some implementations, the specific deuterated ETBR antagonist is a compound of formula (3):

[0130]

[0131] Where X, m, n, R1, R3, R 2a R 2b R4, R5, and R6 are as described in this article.

[0132] In some embodiments, the compound has the following formula (3a):

[0133]

[0134] Where X and R 2a R 2b R4, R5, and R6 are as described in this article.

[0135] In some implementations, the specific deuterated ETBR antagonist is a compound of formula (4):

[0136]

[0137] Where X, m, n, R1, R3, R 2a R 2b R4, R5, and R6 are as described in this article.

[0138] In some embodiments, the compound has the following formula (4a):

[0139]

[0140] Among them, X and R 2a R 2b R4, R5, and R6 are as described in this article.

[0141] In some implementations, the specific deuterated ETBR antagonist is a compound of formula (5):

[0142]

[0143] Where X, m, n, R1, R3, R 2a R 2b R4, R5, and R6 are as described in this article.

[0144] In some embodiments, the compound has the following formula (5a):

[0145]

[0146] Where X and R 2a R 2b R4, R5, and R6 are as described in this article.

[0147] In some implementations, the specific deuterated ETBR antagonist is a compound of formula (6):

[0148]

[0149] Where n, R1, R 2a and R 2b As described in this article.

[0150] In some embodiments, the compound has the following formula (6a):

[0151]

[0153] Where R 2a and R 2b As described in this article.

[0154] In some embodiments of the specific deuterated ETBR antagonists of formulas (1) to (6), n is 0 or 1. In some embodiments, n is 0. In some embodiments, n is 1.

[0155] In some embodiments of the specific deuterated ETBR antagonists of formulas (1) to (5), m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0156] In some embodiments of the specific deuterated ETBR antagonists of formulas (1)-(6), n is 1 and R1 is -D.

[0157] In some embodiments of the specific deuterated ETBR antagonists of formulas (1) to (5a), X is selected from Na + and K + In some implementations, X is Na. + .

[0158] In some embodiments of the specific deuterated ETBR antagonists of formulas (1)-(6a), R 2a and R 2b It is -CH2D.

[0159] In some embodiments of the specific deuterated ETBR antagonists of formulas (1)-(6a), R 2a and R 2b It is -CH3.

[0160] In some embodiments of the specific deuterated ETBR antagonists of formulas (1)-(6a), R 2a or R 2b One of them is CH2D and the other is -CH3.

[0161] In some embodiments, the specific deuterated ETBR antagonist is BQ-788-A, BQ-788-B, BQ-788-C, or a combination thereof, including analogs, derivatives, polymorphs, prodrugs, and salts thereof, including fluorinated analogs. For example, the specific deuterated ETBR antagonist may be a fluorinated analog of BQ-788-A, BQ-788-B, or BQ-788-C.

[0162] In some implementations, the specific deuterated ETBR antagonist is BQ-788-A, as described below: Or its stereoisomers. In some embodiments, the specific deuterated ETBR antagonist is BQ-788-A.

[0163] In some implementations, the specific deuterated ETBR antagonist is BQ-788-B, as described below: Or its stereoisomers. In some embodiments, the specific deuterated ETBR antagonist is BQ-788-B (also known as ENB-003).

[0164] In some implementations, the specific deuterated ETBR antagonist is BQ-788-C, as described below:

[0165] Or its stereoisomers. In some embodiments, the specific deuterated ETBR antagonist is BQ-788-C.

[0166] Pharmaceutical Composition

[0167] In various embodiments, this document provides pharmaceutical compositions comprising an ETBR antagonist (e.g., a specific deuterated ETBR antagonist, such as the deuterated form of BQ-788 as described herein) and a pharmaceutically acceptable carrier.

[0168] In some embodiments, the compositions herein are formulated in unit dosage forms containing any desired carrier or excipient and configured for administration via any desired route, such as oral, intravenous, subcutaneous, intramuscular, intraperitoneal, parenteral, intranasal, or intracranial.

[0169] In some embodiments, the pharmaceutical composition is delivered intravenously, intramuscularly, subcutaneously, orally, intranasally, sublingually, percutaneously, superficially, intraperitoneally, parenterally, intranasally, or intracranially.

[0170] In some implementations, the ETBR antagonist (e.g., a deuterated ETBR antagonist or a deuterated BQ-788) is administered in the form of a liposomal formulation as described herein.

[0171] In some embodiments, the pharmaceutical composition comprises about 1% to about 95% of the active ingredient, a single-dose administration form comprising about 20% to about 90% of the active ingredient, and a non-single-dose administration form comprising about 5% to about 20% of the active ingredient. Unit dosage forms are, for example, coated tablets, tablets, ampoules, vials, suppositories, or capsules. Other administration forms are, for example, ointments, creams, pastes, foams, tinctures, lip balms, drops, sprays, dispersions, etc. An example is a capsule containing about 0.05g to about 5.0g of the active ingredient.

[0172] In some embodiments, the active ingredient is contained in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver a therapeutically effective amount to a patient for the desired indication without causing serious toxicity in the treated patient. Exemplary doses of the active compound used for all conditions mentioned herein range from about 10 ng / kg to 300 mg / kg, for example from 0.001 mg / kg / day to 100 mg / kg / day, and more typically from 0.01 mg / kg recipient / patient weight / day to about 0.5 mg / kg recipient / patient weight / day.

[0173] Typical surface doses will be in the range of 0.01%–5% wt / wt in a suitable carrier. The compound can be conveniently administered in any suitable unit dosage form, including but not limited to unit dosage forms containing the following: 0.01 mg to 50 mg, 0.01 mg to 40 mg, 0.01 mg to 30 mg, 0.01 mg to 20 mg, 0.01 mg to 15 mg, 0.01 mg to 10 mg, 0.1 mg to 50 mg, 0.1 mg to 40 mg, 0.1 mg to 30 mg, 0.1 mg to 20 mg, 0.1 mg to 15 mg, 0.1 mg to 10 mg, 1 mg to 50 mg, 1 mg to 40 mg per unit dosage form. The active ingredient is present in doses of 1 mg to 30 mg, 1 mg to 20 mg, 1 mg to 15 mg, 1 mg to 10 mg, 5 mg to 50 mg, 5 mg to 40 mg, 5 mg to 30 mg, 5 mg to 20 mg, 5 mg to 15 mg, 5 mg to 10 mg, 7.5 mg to 50 mg, 7.5 mg to 40 mg, 7.5 mg to 30 mg, 7.5 mg to 20 mg, 7.5 mg to 15 mg, 7.5 mg to 10 mg, 10 mg to 50 mg, 10 mg to 40 mg, 10 mg to 30 mg, or 10 mg to 20 mg. In some embodiments, the composition comprises an ETBR antagonist (e.g., a specific deuterated ETBR antagonist) in doses of about 0.1 mg to about 50 mg (e.g., about 1 mg to about 25 mg) and / or an ETBR antagonist (e.g., a specific deuterated ETBR antagonist) in concentrations of about 0.001 mg / mL to about 10 mg / mL (e.g., about 0.01 mg / mL to about 1 mg / mL).

[0174] In some embodiments, the compositions described herein are formulated in a conventional manner using one or more pharmaceutically acceptable carriers and may also be administered as controlled-release formulations. Pharmaceutically acceptable carriers that may be used in these pharmaceutical compositions include, but are not limited to, dimethyl sulfoxide (DMSO), soybean oil as a carrier, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of saturated vegetable fatty acids in the form of glycerides, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium dihydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin.

[0175] In some embodiments, the composition comprises soybean oil, dimethyl sulfoxide (DMSO), hydrogel, or a combination thereof. Any embodiment described herein can be a single-component oil-phase formulation as described above, wherein each active ingredient can be any dose or concentration described herein. The single-component oil phase can be a non-volatile oil, such as soybean oil. For example, the formulation contains about 0.1 mg to about 5.0 mg of each active ingredient in 1 mL of single-component oil (i.e., about 0.5 mg / mL, about 1 mg / mL, or about 1.5 mg / mL of each active ingredient in the single-component oil). The single-component oil-phase formulation can be prepared by adding each active ingredient (e.g., about 1 mg to about 50 mg of each active ingredient) to about 10 mL of single-component oil solution.

[0176] In some embodiments, the pharmaceutical compositions herein comprise DMSO, for example, in a DMSO solution of about 5% to about 100% (e.g., about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 90% to about 100%, about 30% to about 95%, about 45% to about 95%, about 75% to about 95%, about 30% to about 90%, about 45% to about 90%, about 75% to about 90%, about 30% to about 85%, about 45% to about 85%, or about 75% to about 85%). For example, a pharmaceutical composition may contain about 0.01 mg to about 5.0 mg of each active ingredient in 1 mL of DMSO (i.e., about 0.05 mg / mL, about 0.1 mg / mL, about 0.15 mg / mL, about 0.2 mg / mL, about 0.3 mg / mL, about 0.5 mg / mL, about 1 mg / mL, or about 1.5 mg / mL of each active ingredient in DMSO). A DMSO pharmaceutical composition may be prepared by adding each active ingredient (e.g., about 0.1 mg to about 50 mg of each active ingredient) to about 10 mL of DMSO solution. For example, DMSO is a DMSO solution containing about 5% to about 100% DMSO, about 25% to about 100% DMSO, about 50% to about 100% DMSO, about 75% to about 100% DMSO, about 5% to about 75% DMSO, about 25% to about 75% DMSO, about 50% to about 75% DMSO, about 5% to about 50% DMSO, about 25% to about 50% DMSO, or about 5% to about 25% DMSO.

[0177] In some embodiments, this specification provides controlled-release subcutaneous or intramuscular dose formulations comprising a homogeneous dispersion of an ETBR antagonist (e.g., a specific deuterated ETBR antagonist, such as BQ-788, its deuterated or fluorinated analogues, or combinations thereof).

[0178] In some embodiments, the pharmaceutical compositions described herein are formulated as controlled-release delivery systems comprising biocompatible polymers. In some embodiments, the active compound is prepared together with a carrier that protects the compound from rapid elimination from the body, such as controlled-release formulations, including implants, hydrogels, thermosensitive hydrogels, and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene-vinyl acetate, acrylates, polycarboxylic acids, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. In some embodiments, the biocompatible polymer is poly(lactide), poly(glycolic acid), poly(lactide-co-glycolic acid), poly(lactic acid), poly(glycolic acid), poly(lactic acid-co-glycolic acid), polycaprolactone, polycarbonate, polyesteramide, polyanhydride, poly(amino acid), polyorthoester, polycyanoacrylate, poly(p-dioxanone), poly(alkylene oxalate), biodegradable polyurethane, blends, or copolymers thereof.

[0179] In some embodiments, the pharmaceutically acceptable carrier comprises or is a liposome. For example, a pharmaceutical composition or formulation may comprise a liposome having an internal volume containing an ETBR antagonist (e.g., a specific deuterated ETBR antagonist). In some embodiments, the liposome is configured to achieve controlled release of the ETBR antagonist (e.g., a specific deuterated ETBR antagonist), such as rapid release, prolonged release, or a combination thereof.

[0180] In some embodiments, liposomes are configured to achieve controlled release of the pharmaceutical composition. In some embodiments, liposomes are configured to achieve rapid release of the pharmaceutical composition. In other embodiments, liposomes are configured or formulated to achieve prolonged release of the pharmaceutical composition. In some embodiments, liposomes are configured to cause both rapid and prolonged release of the pharmaceutical composition.

[0181] In some embodiments, the liposome suspension is a pharmaceutically acceptable carrier. For example, a liposome formulation can be prepared by dissolving one or more suitable lipids (such as stearoylphosphatidylethanolamine, stearoylphosphatidylcholine, arachidonicylphosphatidylcholine, and cholesterol) in an inorganic solvent, followed by evaporation, leaving a dried lipid film on the surface of a container. An aqueous solution of the active compound is then introduced into the container. The container is then manually rotated to move the lipid material away from the container edge and disperse the lipid aggregates, thereby forming a liposome suspension. In some embodiments, the liposomes comprise neutral lipids, basic (with a net positive charge) lipids, acidic (with a net negative charge) lipids, cholesterol, or combinations thereof. In some embodiments, the liposomes also contain a polymer component. In some embodiments, the internal volume of the liposomes is at least partially aqueous and contains an ETBR antagonist (e.g., a specifically deuterated ETBR antagonist).

[0182] In some embodiments, this specification provides pharmaceutical compositions as described herein in liposome delivery systems, such as phosphatidylethanolamine (PE) like dipalmitoyl PE (DPPE), and partially unsaturated phosphatidylcholine (PC) such as ovomucoid PC (EPC) or SPC, fully unsaturated PC such as HSPC, PG, phosphatidylserine (PS), phosphatidylinositol (PI), or combinations thereof. In some embodiments, the phospholipid is a partially unsaturated PG, dipalmitoylphosphatidylglycerol (DPPG), cholesterol, DSPE-PEG2000, polysorbate-80, or combinations thereof. In some embodiments, the liposome delivery system is a controlled-release system, such as at least one or a combination of rapid release, prolonged release, rapid and prolonged release, delayed release, sustained release, and slow release.

[0183] In some embodiments, the pharmaceutical compositions herein comprise pharmaceutically acceptable salts of compounds as described herein, particularly acid or base addition salts. The acids used to prepare pharmaceutically acceptable acid addition salts of the aforementioned base compounds useful according to this aspect are those that form non-toxic acid addition salts, i.e., salts containing pharmaceutically acceptable anions, such as hydrochlorides, hydrobromides, hydroiodides, nitrates, sulfates, hydrogen sulfates, tartrates, phosphates, acid phosphates, acetates, lactates, citrates, acid citrates, tartrates, tartrates, succinates, maleates, fumarates, gluconates, sucrose, benzoates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, and bis(hydroxynaphthyl)ate [i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthylcarbamate)] salts. Pharmaceutically acceptable base addition salts can also be used to prepare pharmaceutically acceptable salt forms of compounds or derivatives according to this disclosure. Chemical bases that can be used as reagents for preparing pharmaceutically acceptable alkali salts of the compounds of the present invention are those that form non-toxic alkali salts with such compounds, which are acidic in nature. Such non-toxic alkali salts include, but are not limited to, those derived from such pharmacologically acceptable cations (such as alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium, zinc, and magnesium), ammonium or water-soluble amine addition salts such as N-methylglucosamine (glucosamine), as well as other alkali salts of lower alkanol ammonium and pharmaceutically acceptable organic amines.

[0184] In some embodiments, the oral composition comprises an inert diluent or an edible carrier. These may be encapsulated in gelatin capsules or compressed into tablets. For oral therapeutic administration, the active compound or its prodrug derivative may be incorporated into excipients and used in tablet, lozenge, or capsule form. Pharmaceutically compatible binders and / or excipients may be included as part of the composition. Tablets, pills, capsules, lozenges, etc., may contain any of the following components or compounds of similar nature: binders, such as microcrystalline cellulose, tragacanth gum, or gelatin; excipients, such as starch or lactose; dispersants, such as alginate, Primogel, or corn starch; lubricants, such as magnesium stearate or sterotes; flow aids, such as colloidal silica; sweeteners, such as sucrose or saccharin; or flavoring agents, such as peppermint, methyl salicylate, or citrus flavorings. When the dosage unit is in capsule form, in addition to the materials of the types described above, the capsule may also contain a liquid carrier such as fatty oil. In addition, the dosage unit form can contain a variety of other materials that change the physical form of the dosage unit, such as coatings of sugar, shellac, or enteric solvents.

[0185] In some embodiments, the active compound or a pharmaceutically acceptable salt thereof is applied as a component of elixirs, suspensions, syrups, rice paper wrappers, chewing gum, etc. In addition to the active compound, the syrup may contain sucrose as a sweetener and certain preservatives, dyes and colorants, and flavorings.

[0186] In some embodiments, solutions or suspensions for parenteral, intradermal, subcutaneous, intravenous, intramuscular, or topical application comprise the following components: sterile diluents such as water for injection, saline solution, non-volatile oils (e.g., soybean oil), polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetate, citrate, or phosphate; and tonic agents such as sodium chloride or dextrose. Parenteral formulations may be encapsulated in ampoules, disposable syringes, or multi-dose vials made of glass or plastic. In some embodiments, the carrier for intravenous administration is physiological saline or phosphate-buffered saline (PBS).

[0187] In some embodiments, each active ingredient of the pharmaceutical composition of this disclosure may be present in any dosage form (e.g., initial burst release, sustained release, etc.) in the following amounts: about 50 μg to about 100,000 μg, about 50 μg to about 90,000 μg, about 50 μg to about 80,000 μg, about 50 μg to about 70,000 μg, about 50 μg to about 60,000 μg, about 50 μg to about 50,000 μg, about 50 μg to about 40,000 μg, about 50 μg to about 30,000 μg, about 50 μg to about 25,000 μg, about 50 μg to about 20,000 μg, about 50 μg to about 15,000 μg, about 50 μg to about 10,000 μg, about 50 μg to about 9000μg, about 50μg to about 8000μg, about 50μg to about 7000μg, about 50μg to about 6000μg, about 50μg to about 5000μg, about 50μg to about 4000μg, about 50μg to about 3000μg, about 50μg to about 2500μg, about 50μg to about 2000μg, about 50μg to about 1500μg, about 50μg to about 1000μg, about 50μg to about 750μg, about 50μg to about 500μg, about 50μg to about 400μg, about 50μg to about 300μg, about 50μg to about 200μg, about 50μg to about 100μg, about 100μg to about 100000μg, about 100μg to about 90,000 μg, about 100 μg to about 80,000 μg, about 100 μg to about 70,000 μg, about 100 μg to about 60,000 μg, about 100 μg to about 50,000 μg, about 100 μg to about 40,000 μg, about 100 μg to about 30,000 μg, about 100 μg to about 25,000 μg, about 100 μg to about 20,000 μg, about 100 μg to about 15,000 μg, about 100 μg to about 10,000 μg, about 100 μg to about 9,000 μg, about 100 μg to about 8,000 μg, about 100 μg to about 7,000 μg, about 100 μg to about 6,000 μg, about 100 μg to about 5,000 μg, about 100 μg to Approximately 4000 μg, approximately 100 μg to approximately 3000 μg, approximately 100 μg to approximately 2500 μg, approximately 100 μg to approximately 2000 μg, approximately 100 μg to approximately 1500 μg, approximately 100 μg to approximately 1000 μg, approximately 100 μg to approximately 750 μg, approximately 100 μg to approximately 500 μg, approximately 100 μg to approximately 400 μg, approximately 100 μg to approximately 300 μg, approximately 100 μg to approximately 200 μg, approximately 200 μg to approximately 100000 μg, approximately 200 μg to approximately 90000 μg, approximately 200 μg to approximately 80000 μg, approximately 200 μg to approximately 70000 μg, approximately 200 μg to approximately 60000 μg, approximately 200 μg to approximately 50000 μg,Approximately 200 μg to approximately 40,000 μg, approximately 200 μg to approximately 30,000 μg, approximately 200 μg to approximately 25,000 μg, approximately 200 μg to approximately 20,000 μg, approximately 200 μg to approximately 15,000 μg, approximately 200 μg to approximately 10,000 μg, approximately 200 μg to approximately 9,000 μg, approximately 200 μg to approximately 8,000 μg, approximately 200 μg to approximately 7,000 μg, approximately 200 μg to approximately 6,000 μg, approximately 200 μg to approximately 5,000 μg, approximately 200 μg to approximately 4,000 μg, approximately 200 μg to approximately 3,000 μg, approximately 200 μg to approximately 2,500 μg, approximately 200 μg to approximately 2,000 μg, approximately 200 μg to approximately 1,500 μg, approximately 200 μg μg to about 1000 μg, about 200 μg to about 750 μg, about 200 μg to about 500 μg, about 200 μg to about 400 μg, about 200 μg to about 300 μg, about 300 μg to about 100000 μg, about 300 μg to about 90000 μg, about 300 μg to about 80000 μg, about 300 μg to about 70000 μg, about 300 μg to about 60000 μg, about 300 μg to about 50000 μg, about 300 μg to about 40000 μg, about 300 μg to about 30000 μg, about 300 μg to about 25000 μg, about 300 μg to about 20000 μg, about 300 μg to about 15000 μg, about 300 μg Approximately 10,000 μg, approximately 300 μg to approximately 9,000 μg, approximately 300 μg to approximately 8,000 μg, approximately 300 μg to approximately 7,000 μg, approximately 300 μg to approximately 6,000 μg, approximately 300 μg to approximately 5,000 μg, approximately 300 μg to approximately 4,000 μg, approximately 300 μg to approximately 3,000 μg, approximately 300 μg to approximately 2,500 μg, approximately 300 μg to approximately 2,000 μg, approximately 300 μg to approximately 1,500 μg, approximately 300 μg to approximately 1,000 μg, approximately 300 μg to approximately 750 μg, approximately 300 μg to approximately 500 μg, approximately 300 μg to approximately 400 μg, approximately 400 μg to approximately 100,000 μg, approximately 400 μg to approximately 90,000 μg. Approximately 400 μg to approximately 80,000 μg, approximately 400 μg to approximately 70,000 μg, approximately 400 μg to approximately 60,000 μg, approximately 400 μg to approximately 50,000 μg, approximately 400 μg to approximately 40,000 μg, approximately 400 μg to approximately 30,000 μg, approximately 400 μg to approximately 25,000 μg, approximately 400 μg to approximately 20,000 μg, approximately 400 μg to approximately 15,000 μg, approximately 400 μg to approximately 10,000 μg, approximately 400 μg to approximately 9,000 μg, approximately 400 μg to approximately 8,000 μg, approximately 400 μg to approximately 7,000 μg, approximately 400 μg to approximately 6,000 μg, approximately 400 μg to approximately 5,000 μg, approximately 400 μg to approximately 4,000 μg,Approximately 400 μg to approximately 3000 μg, approximately 400 μg to approximately 2500 μg, approximately 400 μg to approximately 2000 μg, approximately 400 μg to approximately 1500 μg, approximately 400 μg to approximately 1000 μg, approximately 400 μg to approximately 750 μg, approximately 400 μg to approximately 500 μg, approximately 500 μg to approximately 100000 μg, approximately 500 μg to approximately 90000 μg, approximately 500 μg to approximately 80000 μg, approximately 500 μg to approximately 70000 μg, approximately 500 μg to approximately 60000 μg, approximately 500 μg to approximately 50000 μg, approximately 500 μg to approximately 40000 μg, approximately 500 μg to approximately 30000 μg, approximately 500 μg to approximately 25000 μg g, about 500 μg to about 20000 μg, about 500 μg to about 15000 μg, about 500 μg to about 10000 μg, about 500 μg to about 9000 μg, about 500 μg to about 8000 μg, about 500 μg to about 7000 μg, about 500 μg to about 6000 μg, about 500 μg to about 5000 μg, about 500 μg to about 4000 μg, about 500 μg to about 3000 μg, about 500 μg to about 2500 μg, about 500 μg to about 2000 μg, about 500 μg to about 1500 μg, about 500 μg to about 1000 μg, about 500 μg to about 750 μg, about 750 μg to about 100000 μg, about 750 μg to about 90,000 μg, about 750 μg to about 80,000 μg, about 750 μg to about 70,000 μg, about 750 μg to about 60,000 μg, about 750 μg to about 50,000 μg, about 750 μg to about 40,000 μg, about 750 μg to about 30,000 μg, about 750 μg to about 25,000 μg, about 750 μg to about 20,000 μg, about 750 μg to about 15,000 μg, about 750 μg to about 10,000 μg, about 750 μg to about 9,000 μg, about 750 μg to about 8,000 μg, about 750 μg to about 7,000 μg, about 750 μg to about 6,000 μg, about 750 μg to about 5,000 μg μg, about 750μg to about 4000μg, about 750μg to about 3000μg, about 750μg to about 2500μg, about 750μg to about 2000μg, about 750μg to about 1500μg, about 750μg to about 1000μg, about 1000μg to about 100000μg, about 1000μg to about 90000μg, about 1000μg to about 80000μg, about 1000μg to about 70000μg, about 1000μg to about 60000μg, about 1000μg to about 50000μg, about 1000μg to about 40000μg, about 1000μg to about 30000μg, about 1000μg to about 25000μg,Approximately 1000 μg to approximately 20000 μg, approximately 1000 μg to approximately 15000 μg, approximately 1000 μg to approximately 10000 μg, approximately 1000 μg to approximately 9000 μg, approximately 1000 μg to approximately 8000 μg, approximately 1000 μg to approximately 7000 μg, approximately 1000 μg to approximately 6000 μg, approximately 1000 μg to approximately 5000 μg, approximately 1000 μg to approximately 4000 μg, approximately 1000 μg to approximately 3000 μg, approximately 1000 μg to approximately 2500 μg, approximately 1000 μg to approximately 2000 μg, approximately 1000 μg to approximately 1500 μg, approximately 1500 μg to approximately 100000 μg, approximately 1500 μg to approximately 90000 μg, approximately 15 00 μg to about 80000 μg, about 1500 μg to about 70000 μg, about 1500 μg to about 60000 μg, about 1500 μg to about 50000 μg, about 1500 μg to about 40000 μg, about 1500 μg to about 30000 μg, about 1500 μg to about 25000 μg, about 1500 μg to about 20000 μg, about 1500 μg to about 15000 μg, about 1500 μg to about 10000 μg, about 1500 μg to about 9000 μg, about 1500 μg to about 8000 μg, about 1500 μg to about 7000 μg, about 1500 μg to about 6000 μg, about 1500 μg to about 5000 μg, about 1 500 μg to about 4000 μg, about 1500 μg to about 3000 μg, about 1500 μg to about 2500 μg, about 1500 μg to about 2000 μg, about 2000 μg to about 100000 μg, about 2000 μg to about 90000 μg, about 2000 μg to about 80000 μg, about 2000 μg to about 70000 μg, about 2000 μg to about 60000 μg, about 2000 μg to about 50000 μg, about 2000 μg to about 40000 μg, about 2000 μg to about 30000 μg, about 2000 μg to about 25000 μg, about 2000 μg to about 20000 μg, about 2000 μg to about 15000 μg Approximately 2000 μg to approximately 10000 μg, approximately 2000 μg to approximately 9000 μg, approximately 2000 μg to approximately 8000 μg, approximately 2000 μg to approximately 7000 μg, approximately 2000 μg to approximately 6000 μg, approximately 2000 μg to approximately 5000 μg, approximately 2000 μg to approximately 4000 μg, approximately 2000 μg to approximately 3000 μg, approximately 2000 μg to approximately 2500 μg, approximately 2500 μg to approximately 100000 μg, approximately 2500 μg to approximately 90000 μg, approximately 2500 μg to approximately 80000 μg, approximately 2500 μg to approximately 70000 μg, approximately 2500 μg to approximately 60000 μg, approximately 2500 μg to approximately 50000 μg.Approximately 2500 μg to approximately 40000 μg, approximately 2500 μg to approximately 30000 μg, approximately 2500 μg to approximately 25000 μg, approximately 2500 μg to approximately 20000 μg, approximately 2500 μg to approximately 15000 μg, approximately 2500 μg to approximately 10000 μg, approximately 2500 μg to approximately 9000 μg, approximately 2500 μg to approximately 8000 μg, approximately 2500 μg to approximately 7000 μg, approximately 2500 μg to approximately 6000 μg, approximately 2500 μg to approximately 5000 μg, approximately 2500 μg to approximately 4000 μg, approximately 2500 μg to approximately 3000 μg, approximately 3000 μg to approximately 100000 μg, approximately 3000 μg to approximately 90000 μg g, about 3000μg to about 80000μg, about 3000μg to about 70000μg, about 3000μg to about 60000μg, about 3000μg to about 50000μg, about 3000μg to about 40000μg, about 3000μg to about 30000μg, about 3000μg to about 25000μg, about 3000μg to about 20000μg, about 3000μg to about 15000μg, about 3000μg to about 10000μg, about 3000μg to about 9000μg, about 3000μg to about 8000μg, about 3000μg to about 7000μg, about 3000μg to about 6000μg, about 3000μg to about 50 00 μg, about 3000 μg to about 4000 μg, about 4000 μg to about 100000 μg, about 4000 μg to about 90000 μg, about 4000 μg to about 80000 μg, about 4000 μg to about 70000 μg, about 4000 μg to about 60000 μg, about 4000 μg to about 50000 μg, about 4000 μg to about 40000 μg, about 4000 μg to about 30000 μg, about 4000 μg to about 25000 μg, about 4000 μg to about 20000 μg, about 4000 μg to about 15000 μg, about 4000 μg to about 10000 μg, about 4000 μg to about 9000 μg, about 4000 μg μg to about 8000 μg, about 4000 μg to about 7000 μg, about 4000 μg to about 6000 μg, about 4000 μg to about 5000 μg, about 5000 μg to about 100000 μg, about 5000 μg to about 90000 μg, about 5000 μg to about 80000 μg, about 5000 μg to about 70000 μg, about 5000 μg to about 60000 μg, about 5000 μg to about 50000 μg, about 5000 μg to about 40000 μg, about 5000 μg to about 30000 μg, about 5000 μg to about 25000 μg, about 5000 μg to about 20000 μg, about 5000 μg to about 15000 μg.Approximately 5000 μg to approximately 10000 μg, approximately 5000 μg to approximately 9000 μg, approximately 5000 μg to approximately 8000 μg, approximately 5000 μg to approximately 7000 μg, approximately 5000 μg to approximately 6000 μg, approximately 6000 μg to approximately 100000 μg, approximately 6000 μg to approximately 90000 μg, approximately 6000 μg to approximately 80000 μg, approximately 6000 μg to approximately 70000 μg, approximately 6000 μg to approximately 60000 μg, approximately 6000 μg to approximately 60000 μg, approximately 6000 μg to approximately 50000 μg, approximately 6000 μg to approximately 40000 μg, approximately 6000 μg to approximately 30000 μg, approximately 6000 μg to approximately 25000 μg, approximately 6000 μg to approximately 20000 μg μg, about 6000μg to about 15000μg, about 6000μg to about 10000μg, about 6000μg to about 9000μg, about 6000μg to about 8000μg, about 6000μg to about 7000μg, about 7000μg to about 100000μg, about 7000μg to about 90000μg, about 7000μg to about 80000μg, about 7000μg to about 70000μg, about 7000μg to about 60000μg, about 7000μg to about 50000μg, about 7000μg to about 40000μg, about 7000μg to about 30000μg, about 7000μg to about 25000μg, about 7000μg to about 2 0000μg, about 7000μg to about 15000μg, about 7000μg to about 10000μg, about 7000μg to about 9000μg, about 7000μg to about 8000μg, about 8000μg to about 100000μg, about 8000μg to about 90000μg, about 8000μg to about 80000μg, about 8000μg to about 70000μg, about 8000μg to about 60000μg, about 8000μg to about 50000μg, about 8000μg to about 40000μg, about 8000μg to about 30000μg, about 8000μg to about 25000μg, about 8000μg to about 20000μg, about 8000μg μg to about 15000 μg, about 8000 μg to about 10000 μg, about 8000 μg to about 9000 μg, about 9000 μg to about 100000 μg, about 9000 μg to about 90000 μg, about 9000 μg to about 80000 μg, about 9000 μg to about 70000 μg, about 9000 μg to about 60000 μg, about 9000 μg to about 50000 μg, about 9000 μg to about 40000 μg, about 9000 μg to about 30000 μg, about 9000 μg to about 25000 μg, about 9000 μg to about 20000 μg, about 9000 μg to about 15000 μg, about 9000 μg to about 10000 μg,Approximately 10,000 μg to approximately 100,000 μg, approximately 10,000 μg to approximately 90,000 μg, approximately 10,000 μg to approximately 80,000 μg, approximately 10,000 μg to approximately 70,000 μg, approximately 10,000 μg to approximately 60,000 μg, approximately 10,000 μg to approximately 50,000 μg, approximately 10,000 μg to approximately 40,000 μg, approximately 10,000 μg to approximately 30,000 μg, approximately 10,000 μg to approximately 25,000 μg, approximately 10,000 μg to approximately 20,000 μg, approximately 10,000 μg to approximately 15,000 μg, approximately 15,000 μg to approximately 100,000 μg, approximately 15,000 μg to approximately 90,000 μg, approximately 15,000 μg to approximately 800 μg 00 μg, about 15,000 μg to about 70,000 μg, about 15,000 μg to about 60,000 μg, about 15,000 μg to about 50,000 μg, about 15,000 μg to about 40,000 μg, about 15,000 μg to about 30,000 μg, about 15,000 μg to about 25,000 μg, about 15,000 μg to about 20,000 μg, about 20,000 μg to about 100,000 μg, about 20,000 μg to about 90,000 μg, about 20,000 μg to about 80,000 μg, about 20,000 μg to about 70,000 μg, about 20,000 μg to about 60,000 μg, about 20,000 μg to about 50,000 μg, about 20,000 μg to about 40,000 μg, about 20,000 μg to about 30,000 μg, about 20,000 μg to about 25,000 μg, about 30,000 μg to about 100,000 μg, about 30,000 μg to about 90,000 μg, about 30,000 μg to about 80,000 μg, about 30,000 μg to about 70,000 μg, about 30,000 μg to about 60,000 μg, about 30,000 μg to about 50,000 μg, about 30,000 μg to about 40,000 μg, about 40,000 μg to about 100,000 μg, about 40,000 μg to about 90,000 μg, about 40,000 μg to about 80,000 μg, about 40,000 μg to about 70,000 μg, about 40,000 μg 0 μg to about 60,000 μg, about 40,000 μg to about 50,000 μg, about 50,000 μg to about 100,000 μg, about 50,000 μg to about 90,000 μg, about 50,000 μg to about 80,000 μg, about 50,000 μg to about 70,000 μg, about 50,000 μg to about 60,000 μg, about 60,000 μg to about 100,000 μg, about 60,000 μg to about 90,000 μg, about 60,000 μg to about 80,000 μg, about 60,000 μg to about 70,000 μg, about 70,000 μg to about 100,000 μg, about 70,000 μg to about 90,000 μg, about 70,000 μg to about 80,000 μgApproximately 80,000 μg to approximately 100,000 μg, approximately 80,000 μg to approximately 90,000 μg, or approximately 90,000 μg to approximately 100,000 μg.

[0188] In some embodiments, each active ingredient of the pharmaceutical composition of this disclosure may be present in any dosage form (e.g., initial burst release, sustained release, etc.) in the following amounts: about 50 μg to about 500 μg, about 500 μg to about 2000 μg, about 2000 μg to about 10000 μg, about 10000 μg to about 20000 μg, about 20000 μg to about 50000 μg, or about 50000 μg to about 100000 μg.

[0189] In some embodiments, each active ingredient of the pharmaceutical composition of this disclosure is present in the following amounts: about 0.1 mg / mL to about 50 mg / mL, about 0.1 mg / mL to about 25 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 1 mg / mL to about 50 mg / mL, about 1 mg / mL to about 25 mg / mL, about 1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 5.0 mg / mL (e.g., relative to the pharmaceutical composition, about 0.1 mg / mL to about 4.5 mg / mL, about 0.1 mg / mL to about 4.0 mg / mL, about 0.1 mg / mL to about 3.5 mg / mL, about 0.1 mg / mL to about 3.0 mg / mL). / mL, about 0.1 mg / mL to about 2.5 mg / mL, about 0.1 mg / mL to about 2.0 mg / mL, about 0.1 mg / mL to about 1.5 mg / mL, about 0.1 mg / mL to about 1.0 mg / mL, about 0.1 mg / mL to about 0.5 mg / mL, about 0.5 mg / mL to about 4.5 mg / mL, about 0.5 mg / mL to about 4.0 mg / mL, about 0.5 mg / mL to about 3.5 mg / mL, about 0.5 mg / mL to about 3.0 mg / mL, about 0.5 mg / mL to about 2.5 mg / mL, about 0.5 mg / mL to about 2.0 mg / mL, about 0.5 mg / mL to about 1.5 mg / mL, about 0.5 mg / mL g / mL to about 1.0 mg / mL, about 1.0 mg / mL to about 4.5 mg / mL, about 1.0 mg / mL to about 4.0 mg / mL, about 1.0 mg / mL to about 3.5 mg / mL, about 1.0 mg / mL to about 3.0 mg / mL, about 1.0 mg / mL to about 2.5 mg / mL, about 1.0 mg / mL to about 2.0 mg / mL, about 1.0 mg / mL to about 1.5 mg / mL, about 1.5 mg / mL to about 4.5 mg / mL, about 1.5 mg / mL to about 4.0 mg / mL, about 1.5 mg / mL to about 3.5 mg / mL, about 1.5 mg / mL to about 3.0 mg / mL, about 1.5 mg / mL to about 2.5 mg / mL. mg / mL, about 1.5 mg / mL to about 2.0 mg / mL, about 2.0 mg / mL to about 4.5 mg / mL, about 2.0 mg / mL to about 4.0 mg / mL, about 2.0 mg / mL to about 3.5 mg / mL, about 2.0 mg / mL to about 3.0 mg / mL, about 2.0 mg / mL to about 2.5 mg / mL, about 2.5 mg / mL to about 4.5 mg / mL, about 2.5 mg / mL to about 4.0 mg / mL, about 2.5 mg / mL to about 3.5 mg / mL, about 2.5 mg / mL to about 3.0 mg / mL, about 3.0 mg / mL to about 4.5 mg / mL, about 3.0 mg / mL to about 4.0 mg / mL, about 3.(0 mg / mL to about 3.5 mg / mL, about 3.5 mg / mL to about 4.5 mg / mL, about 3.5 mg / mL to about 4.0 mg / mL, or about 3.5 mg / mL to about 4.5 mg / mL).

[0190] In some embodiments, each active ingredient of the pharmaceutical composition of this disclosure is present in amounts of: about 0.1 μg / mL to about 50 μg / mL, about 0.1 μg / mL to about 25 μg / mL, about 0.1 μg / mL to about 10 μg / mL, about 1 μg / mL to about 50 μg / mL, about 1 μg / mL to about 25 μg / mL, about 1 μg / mL to about 10 μg / mL, about 0.1 μg / mL to about 5.0 μg / mL, for example, about 1 μg / mL to about 5 μg / mL, about 0.1 μg / mL to about 4.0 μg / mL, about 0.1 μg / mL to about 3.5 μg / mL, about 0.1 μg / mL to about 3.0 μg / mL, about 0 0.1 μg / mL to about 2.5 μg / mL, about 0.1 μg / mL to about 2.0 μg / mL, about 0.1 μg / mL to about 1.5 μg / mL, about 0.1 μg / mL to about 1.0 μg / mL, about 0.1 μg / mL to about 0.5 μg / mL, about 0.5 μg / mL to about 4.5 μg / mL, about 0.5 μg / mL to about 4.0 μg / mL, about 0.5 μg / mL to about 3.5 μg / mL, about 0.5 μg / mL to about 3.0 μg / mL, about 0.5 μg / mL to about 2.5 μg / mL, about 0.5 μg / mL to about 2.0 μg / mL, about 0.5 μg / mL to about 1.5 μg / mL, about 0.5 μg / mL to about 1 0.0 μg / mL, about 1.0 μg / mL to about 4.5 μg / mL, about 1.0 μg / mL to about 4.0 μg / mL, about 1.0 μg / mL to about 3.5 μg / mL, about 1.0 μg / mL to about 3.0 μg / mL, about 1.0 μg / mL to about 2.5 μg / mL, about 1.0 μg / mL to about 2.0 μg / mL, about 1.0 μg / mL to about 1.5 μg / mL, about 1.5 μg / mL to about 4.5 μg / mL, about 1.5 μg / mL to about 4.0 μg / mL, about 1.5 μg / mL to about 3.5 μg / mL, about 1.5 μg / mL to about 3.0 μg / mL, about 1.5 μg / mL to about 2.5 μg / mL, about 1 0.5 μg / mL to about 2.0 μg / mL, about 2.0 μg / mL to about 4.5 μg / mL, about 2.0 μg / mL to about 4.0 μg / mL, about 2.0 μg / mL to about 3.5 μg / mL, about 2.0 μg / mL to about 3.0 μg / mL, about 2.0 μg / mL to about 2.5 μg / mL, about 2.5 μg / mL to about 4.5 μg / mL, about 2.5 μg / mL to about 4.0 μg / mL, about 2.5 μg / mL to about 3.5 μg / mL, about 2.5 μg / mL to about 3.0 μg / mL, about 3.0 μg / mL to about 4.5 μg / mL, about 3.0 μg / mL to about 4.0 μg / mL, about 3.0 μg / mL to about 3.0 μg / mL.5 μg / mL, about 3.5 μg / mL to about 4.5 μg / mL, about 3.5 μg / mL to about 4.0 μg / mL, or about 3.5 μg / mL to about 4.5 μg / mL.

[0191] Dosage regimen

[0192] Treatment regimens are disclosed herein in various embodiments. In some embodiments, the treatment regimen comprises a dosage pharmaceutical composition having approximately 100 μg to approximately 100,000 μg of each contained active ingredient (i.e., an ETBR antagonist (e.g., a specific deuterated ETBR antagonist as described herein)). The dosage may be a sustained-release dosage, wherein approximately 50 μg to approximately 60,000 μg of each active ingredient is initially released, while approximately 50 μg to approximately 60,000 μg of each active ingredient is sustained-released over 2 hours.

[0193] In some implementations, the dosage of the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) may be from about 1.0 μg to about 100,000 μg.

[0194] In some embodiments, the dosage of the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) may be from about 1.0 μg to about 100,000 μg, from about 1.0 μg to about 90,000 μg, from about 1.0 μg to about 80,000 μg, from about 1.0 μg to about 70,000 μg, from about 1.0 μg to about 60,000 μg, from about 1.0 μg to about 50,000 μg, from about 1.0 μg to about 40,000 μg, from about 1.0 μg to about 30,000 μg, from about 1.0 μg to about 29,000 μg, from about 1.0 μg to about 28,000 μg, from about 1.0 μg to about 27,000 μg, from about 1.0 μg to about 26,000 μg, or from about 1.0 μg to about 250 μg. 0.0 μg, about 1.0 μg to about 24000 μg, about 1.0 μg to about 23000 μg, about 1.0 μg to about 22000 μg, about 1.0 μg to about 21000 μg, about 1.0 μg to about 20000 μg, about 1.0 μg to about 19000 μg, about 1.0 μg to about 18000 μg, about 1.0 μg to about 17000 μg, about 1.0 μg to about 16000 μg, about 1.0 μg to about 15000 μg, about 1.0 μg to about 14000 μg, about 1.0 μg to about 13000 μg, about 1.0 μg to about 12000 μg, about 1.0 μg to about 11000 μg, about 1.0 μg to about 10000 μg, about 1 0 μg to about 9000 μg, about 1.0 μg to about 8000 μg, about 1.0 μg to about 7000 μg, about 1.0 μg to about 6000 μg, about 1.0 μg to about 5000 μg, about 1.0 μg to about 4500 μg, about 1.0 μg to about 4000 μg, about 1.0 μg to about 3500 μg, about 1.0 μg to about 3000 μg, about 1.0 μg to about 2500 μg, about 1.0 μg to about 2000 μg, about 1.0 μg to about 1500 μg, about 1.0 μg to about 1250 μg, about 1.0 μg to about 1000 μg, about 1.0 μg to about 750 μg, about 1.0 μg to about 500 μg, about 1.0 μg to about 300 μg μg, about 1.0 μg to about 250 μg, about 1.0 μg to about 200 μg, about 1.0 μg to about 150 μg, about 1.0 μg to about 100 μg, about 1.0 μg to about 75 μg, about 1.0 μg to about 50 μg, about 1.0 μg to about 25 μg, about 1.0 μg to about 10 μg, about 10 μg to about 100,000 μg, about 10 μg to about 90,000 μg, about 10 μg to about 80,000 μg, about 10 μg to about 70,000 μg, about 10 μg to about 60,000 μg, about 10 μg to about 50,000 μg, about 10 μg to about 40,000 μg, about 10 μg to about 30,000 μg, about 10 μg to about 29,000 μg.About 10 μg to about 28000 μg, about 10 μg to about 27000 μg, about 10 μg to about 26000 μg, about 10 μg to about 25000 μg, about 10 μg to about 24000 μg, about 10 μg to about 23000 μg, about 10 μg to about 22000 μg, about 10 μg to about 21000 μg, about 10 μg to about 20000 μg, about 10 μg to about 19000 μg, about 10 μg to about 18000 μg, about 10 μg to about 17000 μg, about 10 μg to about 16000 μg, about 10 μg to about 15000 μg, about 10 μg to about 14000 μg, about 10 μg to about 13000 μg, about 10 μg to about 12 000μg, about 10μg to about 11000μg, about 10μg to about 10000μg, about 10μg to about 9000μg, about 10μg to about 8000μg, about 10μg to about 7000μg, about 10μg to about 6000μg, about 10μg to about 5000μg, about 10μg to about 4500μg, about 10μg to about 4000μg, about 10μg to about 3500μg, about 10μg to about 3000μg, about 10μg to about 2500μg, about 10μg to about 2000μg, about 10μg to about 1500μg, about 10μg to about 1250μg, about 10μg to about 1000μg, about 10μg to about 750μg, about 10μg About 500 μg, about 10 μg to about 300 μg, about 10 μg to about 250 μg, about 10 μg to about 200 μg, about 10 μg to about 150 μg, about 10 μg to about 100 μg, about 10 μg to about 75 μg, about 10 μg to about 50 μg, about 10 μg to about 25 μg, about 25 μg to about 100,000 μg, about 25 μg to about 90,000 μg, about 25 μg to about 80,000 μg, about 25 μg to about 70,000 μg, about 25 μg to about 60,000 μg, about 25 μg to about 50,000 μg, about 25 μg to about 40,000 μg, about 25 μg to about 30,000 μg, about 25 μg to about 29,000 μg, about 25 μg Approximately 28,000 μg, approximately 25 μg to approximately 27,000 μg, approximately 25 μg to approximately 26,000 μg, approximately 25 μg to approximately 25,000 μg, approximately 25 μg to approximately 24,000 μg, approximately 25 μg to approximately 23,000 μg, approximately 25 μg to approximately 22,000 μg, approximately 25 μg to approximately 21,000 μg, approximately 25 μg to approximately 20,000 μg, approximately 25 μg to approximately 19,000 μg, approximately 25 μg to approximately 18,000 μg, approximately 25 μg to approximately 17,000 μg, approximately 25 μg to approximately 16,000 μg, approximately 25 μg to approximately 15,000 μg, approximately 25 μg to approximately 14,000 μg, approximately 25 μg to approximately 13,000 μg, approximately 25 μg to approximately 12,000 μg.Approximately 25 μg to approximately 11000 μg, approximately 25 μg to approximately 10000 μg, approximately 25 μg to approximately 9000 μg, approximately 25 μg to approximately 8000 μg, approximately 25 μg to approximately 7000 μg, approximately 25 μg to approximately 6000 μg, approximately 25 μg to approximately 5000 μg, approximately 25 μg to approximately 4500 μg, approximately 25 μg to approximately 4000 μg, approximately 25 μg to approximately 3500 μg, approximately 25 μg to approximately 3000 μg, approximately 25 μg to approximately 2500 μg, approximately 25 μg to approximately 2000 μg, approximately 25 μg to approximately 1500 μg, approximately 25 μg to approximately 1250 μg, approximately 25 μg to approximately 1000 μg, approximately 25 μg to approximately 750 μg, approximately 25 μg to approximately 500 μg μg, about 25μg to about 300μg, about 25μg to about 250μg, about 25μg to about 200μg, about 25μg to about 150μg, about 25μg to about 100μg, about 25μg to about 75μg, about 25μg to about 50μg, about 50μg to about 100000μg, about 50μg to about 90000μg, about 50μg to about 80000μg, about 50μg to about 70000μg, about 50μg to about 60000μg, about 50μg to about 50000μg, about 50μg to about 40000μg, about 50μg to about 30000μg, about 50μg to about 29000μg, about 50μg to about 28000μg, about 50μg to Approximately 27,000 μg, approximately 50 μg to approximately 26,000 μg, approximately 50 μg to approximately 25,000 μg, approximately 50 μg to approximately 24,000 μg, approximately 50 μg to approximately 23,000 μg, approximately 50 μg to approximately 22,000 μg, approximately 50 μg to approximately 21,000 μg, approximately 50 μg to approximately 20,000 μg, approximately 50 μg to approximately 19,000 μg, approximately 50 μg to approximately 18,000 μg, approximately 50 μg to approximately 17,000 μg, approximately 50 μg to approximately 16,000 μg, approximately 50 μg to approximately 15,000 μg, approximately 50 μg to approximately 14,000 μg, approximately 50 μg to approximately 13,000 μg, approximately 50 μg to approximately 12,000 μg, approximately 50 μg to approximately 11,000 μg. Approximately 50 μg to approximately 10,000 μg, approximately 50 μg to approximately 9,000 μg, approximately 50 μg to approximately 8,000 μg, approximately 50 μg to approximately 7,000 μg, approximately 50 μg to approximately 6,000 μg, approximately 50 μg to approximately 5,000 μg, approximately 50 μg to approximately 4,500 μg, approximately 50 μg to approximately 4,000 μg, approximately 50 μg to approximately 3,500 μg, approximately 50 μg to approximately 3,000 μg, approximately 50 μg to approximately 2,500 μg, approximately 50 μg to approximately 2,000 μg, approximately 50 μg to approximately 1,500 μg, approximately 50 μg to approximately 1,250 μg, approximately 50 μg to approximately 1,000 μg, approximately 50 μg to approximately 750 μg, approximately 50 μg to approximately 500 μg, approximately 50 μg to approximately 300 μg,Approximately 50 μg to approximately 250 μg, approximately 50 μg to approximately 200 μg, approximately 50 μg to approximately 150 μg, approximately 50 μg to approximately 100 μg, approximately 50 μg to approximately 75 μg, approximately 75 μg to approximately 100,000 μg, approximately 75 μg to approximately 90,000 μg, approximately 75 μg to approximately 80,000 μg, approximately 75 μg to approximately 70,000 μg, approximately 75 μg to approximately 60,000 μg, approximately 75 μg to approximately 50,000 μg, approximately 75 μg to approximately 40,000 μg, approximately 75 μg to approximately 30,000 μg, approximately 75 μg to approximately 29,000 μg, approximately 75 μg to approximately 28,000 μg, approximately 75 μg to approximately 27,000 μg, approximately 75 μg to approximately 26,000 μg, approximately 75 μg Approximately 25,000 μg, approximately 75 μg to approximately 24,000 μg, approximately 75 μg to approximately 23,000 μg, approximately 75 μg to approximately 22,000 μg, approximately 75 μg to approximately 21,000 μg, approximately 75 μg to approximately 20,000 μg, approximately 75 μg to approximately 19,000 μg, approximately 75 μg to approximately 18,000 μg, approximately 75 μg to approximately 17,000 μg, approximately 75 μg to approximately 16,000 μg, approximately 75 μg to approximately 15,000 μg, approximately 75 μg to approximately 14,000 μg, approximately 75 μg to approximately 13,000 μg, approximately 75 μg to approximately 12,000 μg, approximately 75 μg to approximately 11,000 μg, approximately 75 μg to approximately 10,000 μg, approximately 75 μg to approximately 9,000 μg Approximately 75 μg to approximately 8000 μg, approximately 75 μg to approximately 7000 μg, approximately 75 μg to approximately 6000 μg, approximately 75 μg to approximately 5000 μg, approximately 75 μg to approximately 4500 μg, approximately 75 μg to approximately 4000 μg, approximately 75 μg to approximately 3500 μg, approximately 75 μg to approximately 3000 μg, approximately 75 μg to approximately 2500 μg, approximately 75 μg to approximately 2000 μg, approximately 75 μg to approximately 1500 μg, approximately 75 μg to approximately 1250 μg, approximately 75 μg to approximately 1000 μg, approximately 75 μg to approximately 750 μg, approximately 75 μg to approximately 500 μg, approximately 75 μg to approximately 300 μg, approximately 75 μg to approximately 250 μg, approximately 75 μg to approximately 200 μg, approximately 75 μg to about 150 μg, about 75 μg to about 100 μg, about 100 μg to about 100,000 μg, about 100 μg to about 90,000 μg, about 100 μg to about 80,000 μg, about 100 μg to about 70,000 μg, about 100 μg to about 60,000 μg, about 100 μg to about 50,000 μg, about 100 μg to about 40,000 μg, about 100 μg to about 30,000 μg, about 100 μg to about 29,000 μg, about 100 μg to about 28,000 μg, about 100 μg to about 27,000 μg, about 100 μg to about 26,000 μg, about 100 μg to about 25,000 μg, about 100 μg to about 24,000 μg.About 100 μg to about 23000 μg, about 100 μg to about 22000 μg, about 100 μg to about 21000 μg, about 100 μg to about 20000 μg, about 100 μg to about 19000 μg, about 100 μg to about 18000 μg, about 100 μg to about 17000 μg, about 100 μg to about 16000 μg, about 100 μg to about 15000 μg, about 100 μg to about 14000 μg, about 100 μg to about 13000 μg, about 100 μg to about 12000 μg, about 100 μg to about 11000 μg, about 100 μg to about 10000 μg, about 100 μg to about 9000 μg, about 100 μg to about 80 0.00 μg, about 100 μg to about 7000 μg, about 100 μg to about 6000 μg, about 100 μg to about 5000 μg, about 100 μg to about 4500 μg, about 100 μg to about 4000 μg, about 100 μg to about 3500 μg, about 100 μg to about 3000 μg, about 100 μg to about 2500 μg, about 100 μg to about 2000 μg, about 100 μg to about 1500 μg, about 100 μg to about 1250 μg, about 100 μg to about 1000 μg, about 100 μg to about 750 μg, about 100 μg to about 500 μg, about 100 μg to about 300 μg, about 100 μg to about 250 μg, about 100 μg to about 200 μg, about 100 μg to about 150 μg, about 150 μg to about 100,000 μg, about 150 μg to about 90,000 μg, about 150 μg to about 80,000 μg, about 150 μg to about 70,000 μg, about 150 μg to about 60,000 μg, about 150 μg to about 50,000 μg, about 150 μg to about 40,000 μg, about 150 μg to about 30,000 μg, about 150 μg to about 29,000 μg, about 150 μg to about 28,000 μg, about 150 μg to about 27,000 μg, about 150 μg to about 26,000 μg, about 150 μg to about 25,000 μg, about 150 μg to about 24,000 μg, about 150 μg Approximately 23,000 μg to 23,000 μg, approximately 150 μg to 22,000 μg, approximately 150 μg to 21,000 μg, approximately 150 μg to 20,000 μg, approximately 150 μg to 19,000 μg, approximately 150 μg to 18,000 μg, approximately 150 μg to 17,000 μg, approximately 150 μg to 16,000 μg, approximately 150 μg to 15,000 μg, approximately 150 μg to 14,000 μg, approximately 150 μg to 13,000 μg, approximately 150 μg to 12,000 μg, approximately 150 μg to 11,000 μg, approximately 150 μg to 10,000 μg, approximately 150 μg to 9,000 μg, approximately 150 μg to 8,000 μg.Approximately 150 μg to approximately 7000 μg, approximately 150 μg to approximately 6000 μg, approximately 150 μg to approximately 5000 μg, approximately 150 μg to approximately 4500 μg, approximately 150 μg to approximately 4000 μg, approximately 150 μg to approximately 3500 μg, approximately 150 μg to approximately 3000 μg, approximately 150 μg to approximately 2500 μg, approximately 150 μg to approximately 2000 μg, approximately 150 μg to approximately 1500 μg, approximately 150 μg to approximately 1250 μg, approximately 150 μg to approximately 1000 μg, approximately 150 μg to approximately 750 μg, approximately 150 μg to approximately 500 μg, approximately 150 μg to approximately 300 μg, approximately 150 μg to approximately 250 μg, approximately 150 μg to approximately 200 μg, Approximately 250 μg to approximately 100,000 μg, approximately 250 μg to approximately 90,000 μg, approximately 250 μg to approximately 80,000 μg, approximately 250 μg to approximately 70,000 μg, approximately 250 μg to approximately 60,000 μg, approximately 250 μg to approximately 50,000 μg, approximately 250 μg to approximately 40,000 μg, approximately 250 μg to approximately 30,000 μg, approximately 250 μg to approximately 29,000 μg, approximately 250 μg to approximately 28,000 μg, approximately 250 μg to approximately 27,000 μg, approximately 250 μg to approximately 26,000 μg, approximately 250 μg to approximately 25,000 μg, approximately 250 μg to approximately 24,000 μg, approximately 250 μg to approximately 23,000 μg, approximately 250 μg to approximately 2 2000 μg, about 250 μg to about 21000 μg, about 250 μg to about 20000 μg, about 250 μg to about 19000 μg, about 250 μg to about 18000 μg, about 250 μg to about 17000 μg, about 250 μg to about 16000 μg, about 250 μg to about 15000 μg, about 250 μg to about 14000 μg, about 250 μg to about 13000 μg, about 250 μg to about 12000 μg, about 250 μg to about 11000 μg, about 250 μg to about 10000 μg, about 250 μg to about 9000 μg, about 250 μg to about 8000 μg, about 250 μg to about 7000 μg, about 250 μg Approximately 6000 μg, approximately 250 μg to approximately 5000 μg, approximately 250 μg to approximately 4500 μg, approximately 250 μg to approximately 4000 μg, approximately 250 μg to approximately 3500 μg, approximately 250 μg to approximately 3000 μg, approximately 250 μg to approximately 2500 μg, approximately 250 μg to approximately 2000 μg, approximately 250 μg to approximately 1500 μg, approximately 250 μg to approximately 1250 μg, approximately 250 μg to approximately 1000 μg, approximately 250 μg to approximately 750 μg, approximately 250 μg to approximately 500 μg, approximately 250 μg to approximately 300 μg, approximately 300 μg to approximately 100000 μg, approximately 300 μg to approximately 90000 μg, approximately 300 μg to approximately 80000 μg.Approximately 300 μg to approximately 70,000 μg, approximately 300 μg to approximately 60,000 μg, approximately 300 μg to approximately 50,000 μg, approximately 300 μg to approximately 40,000 μg, approximately 300 μg to approximately 30,000 μg, approximately 300 μg to approximately 29,000 μg, approximately 300 μg to approximately 28,000 μg, approximately 300 μg to approximately 27,000 μg, approximately 300 μg to approximately 26,000 μg, approximately 300 μg to approximately 25,000 μg, approximately 300 μg to approximately 24,000 μg, approximately 300 μg to approximately 23,000 μg, approximately 300 μg to approximately 22,000 μg, approximately 300 μg to approximately 21,000 μg, approximately 300 μg to approximately 20,000 μg, approximately 300 μg to Approximately 19,000 μg, approximately 300 μg to approximately 18,000 μg, approximately 300 μg to approximately 17,000 μg, approximately 300 μg to approximately 16,000 μg, approximately 300 μg to approximately 15,000 μg, approximately 300 μg to approximately 14,000 μg, approximately 300 μg to approximately 13,000 μg, approximately 300 μg to approximately 12,000 μg, approximately 300 μg to approximately 11,000 μg, approximately 300 μg to approximately 10,000 μg, approximately 300 μg to approximately 9,000 μg, approximately 300 μg to approximately 8,000 μg, approximately 300 μg to approximately 7,000 μg, approximately 300 μg to approximately 6,000 μg, approximately 300 μg to approximately 5,000 μg, approximately 300 μg to approximately 4,500 μg, approximately 300 μg g to about 4000 μg, about 300 μg to about 3500 μg, about 300 μg to about 3000 μg, about 300 μg to about 2500 μg, about 300 μg to about 2000 μg, about 300 μg to about 1500 μg, about 300 μg to about 1250 μg, about 300 μg to about 1000 μg, about 300 μg to about 750 μg, about 300 μg to about 500 μg, about 500 μg to about 100000 μg, about 500 μg to about 90000 μg, about 500 μg to about 80000 μg, about 500 μg to about 70000 μg, about 500 μg to about 60000 μg, about 500 μg to about 50000 μg, about 500 μg to about 50000 μg, about 500 μg to about 40,000 μg, about 500 μg to about 30,000 μg, about 500 μg to about 29,000 μg, about 500 μg to about 28,000 μg, about 500 μg to about 27,000 μg, about 500 μg to about 26,000 μg, about 500 μg to about 25,000 μg, about 500 μg to about 24,000 μg, about 500 μg to about 23,000 μg, about 500 μg to about 22,000 μg, about 500 μg to about 21,000 μg, about 500 μg to about 20,000 μg, about 500 μg to about 19,000 μg, about 500 μg to about 18,000 μg, about 500 μg to about 17,000 μg, about 500 μg to about 16,000 μg.Approximately 500 μg to approximately 15000 μg, approximately 500 μg to approximately 14000 μg, approximately 500 μg to approximately 13000 μg, approximately 500 μg to approximately 12000 μg, approximately 500 μg to approximately 11000 μg, approximately 500 μg to approximately 10000 μg, approximately 500 μg to approximately 9000 μg, approximately 500 μg to approximately 8000 μg, approximately 500 μg to approximately 7000 μg, approximately 500 μg to approximately 6000 μg, approximately 500 μg to approximately 5000 μg, approximately 500 μg to approximately 4500 μg, approximately 500 μg to approximately 4000 μg, approximately 500 μg to approximately 3500 μg, approximately 500 μg to approximately 3000 μg, approximately 500 μg to approximately 2500 μg, approximately 50 0 μg to about 2000 μg, about 500 μg to about 1500 μg, about 500 μg to about 1250 μg, about 500 μg to about 1000 μg, about 500 μg to about 750 μg, about 750 μg to about 100000 μg, about 750 μg to about 90000 μg, about 750 μg to about 80000 μg, about 750 μg to about 70000 μg, about 750 μg to about 60000 μg, about 750 μg to about 50000 μg, about 750 μg to about 40000 μg, about 750 μg to about 30000 μg, about 750 μg to about 29000 μg, about 750 μg to about 28000 μg, about 750 μg to about 27000 μg, about 750 μg to about 26000 μg, about 750 μg to about 25000 μg, about 750 μg to about 24000 μg, about 750 μg to about 23000 μg, about 750 μg to about 22000 μg, about 750 μg to about 21000 μg, about 750 μg to about 20000 μg, about 750 μg to about 19000 μg, about 750 μg to about 18000 μg, about 750 μg to about 17000 μg, about 750 μg to about 16000 μg, about 750 μg to about 15000 μg, about 750 μg to about 14000 μg, about 750 μg to about 13000 μg, about 750 μg to about 12000 μg, about 750 μg to about 11 000μg, about 750μg to about 10000μg, about 750μg to about 9000μg, about 750μg to about 8000μg, about 750μg to about 7000μg, about 750μg to about 6000μg, about 750μg to about 5000μg, about 750μg to about 4500μg, about 750μg to about 4000μg, about 750μg to about 3500μg, about 750μg to about 3000μg, about 750μg to about 2500μg, about 750μg to about 2000μg, about 750μg to about 1500μg, about 750μg to about 1250μg, about 750μg to about 1000μg, about 1000μg to about 100000μg,Approximately 1000 μg to approximately 90000 μg, approximately 1000 μg to approximately 80000 μg, approximately 1000 μg to approximately 70000 μg, approximately 1000 μg to approximately 60000 μg, approximately 1000 μg to approximately 50000 μg, approximately 1000 μg to approximately 40000 μg, approximately 1000 μg to approximately 30000 μg, approximately 1000 μg to approximately 29000 μg, approximately 1000 μg to approximately 28000 μg, approximately 1000 μg to approximately 27000 μg, approximately 1000 μg to approximately 26000 μg, approximately 1000 μg to approximately 25000 μg, approximately 1000 μg to approximately 24000 μg, approximately 1000 μg to approximately 23000 μg, approximately 1000 μg to approximately 22000 μg, about 1000 μg to about 21000 μg, about 1000 μg to about 20000 μg, about 1000 μg to about 19000 μg, about 1000 μg to about 18000 μg, about 1000 μg to about 17000 μg, about 1000 μg to about 16000 μg, about 1000 μg to about 15000 μg, about 1000 μg to about 14000 μg, about 1000 μg to about 13000 μg, about 1000 μg to about 12000 μg, about 1000 μg to about 11000 μg, about 1000 μg to about 10000 μg, about 1000 μg to about 9000 μg, about 1000 μg to about 8000 μg, about 100 0 μg to about 7000 μg, about 1000 μg to about 6000 μg, about 1000 μg to about 5000 μg, about 1000 μg to about 4500 μg, about 1000 μg to about 4000 μg, about 1000 μg to about 3500 μg, about 1000 μg to about 3000 μg, about 1000 μg to about 2500 μg, about 1000 μg to about 2000 μg, about 1000 μg to about 1500 μg, about 1000 μg to about 1250 μg, about 1250 μg to about 100000 μg, about 1250 μg to about 90000 μg, about 1250 μg to about 80000 μg, about 1250 μg to about 70000 μg, about 1250 μg Approximately 60,000 μg, approximately 1,250 μg to approximately 50,000 μg, approximately 1,250 μg to approximately 40,000 μg, approximately 1,250 μg to approximately 30,000 μg, approximately 1,250 μg to approximately 29,000 μg, approximately 1,250 μg to approximately 28,000 μg, approximately 1,250 μg to approximately 27,000 μg, approximately 1,250 μg to approximately 26,000 μg, approximately 1,250 μg to approximately 25,000 μg, approximately 1,250 μg to approximately 24,000 μg, approximately 1,250 μg to approximately 23,000 μg, approximately 1,250 μg to approximately 22,000 μg, approximately 1,250 μg to approximately 21,000 μg, approximately 1,250 μg to approximately 20,000 μg, approximately 1,250 μg to approximately 19,000 μg.Approximately 1250 μg to approximately 18000 μg, approximately 1250 μg to approximately 17000 μg, approximately 1250 μg to approximately 16000 μg, approximately 1250 μg to approximately 15000 μg, approximately 1250 μg to approximately 14000 μg, approximately 1250 μg to approximately 13000 μg, approximately 1250 μg to approximately 12000 μg, approximately 1250 μg to approximately 11000 μg, approximately 1250 μg to approximately 10000 μg, approximately 1250 μg to approximately 9000 μg, approximately 1250 μg to approximately 8000 μg, approximately 1250 μg to approximately 7000 μg, approximately 1250 μg to approximately 6000 μg, approximately 1250 μg to approximately 5000 μg, approximately 1250 μg to approximately 4500 μg. Approximately 1250 μg to approximately 4000 μg, approximately 1250 μg to approximately 3500 μg, approximately 1250 μg to approximately 3000 μg, approximately 1250 μg to approximately 2500 μg, approximately 1250 μg to approximately 2000 μg, approximately 1250 μg to approximately 1500 μg, approximately 1500 μg to approximately 100000 μg, approximately 1500 μg to approximately 90000 μg, approximately 1500 μg to approximately 80000 μg, approximately 1500 μg to approximately 70000 μg, approximately 1500 μg to approximately 60000 μg, approximately 1500 μg to approximately 50000 μg, approximately 1500 μg to approximately 40000 μg, approximately 1500 μg to approximately 30000 μg, approximately 1500 μg to approximately 29000 μg. Approximately 1500 μg to approximately 28000 μg, approximately 1500 μg to approximately 27000 μg, approximately 1500 μg to approximately 26000 μg, approximately 1500 μg to approximately 25000 μg, approximately 1500 μg to approximately 24000 μg, approximately 1500 μg to approximately 23000 μg, approximately 1500 μg to approximately 22000 μg, approximately 1500 μg to approximately 21000 μg, approximately 1500 μg to approximately 20000 μg, approximately 1500 μg to approximately 19000 μg, approximately 1500 μg to approximately 18000 μg, approximately 1500 μg to approximately 17000 μg, approximately 1500 μg to approximately 16000 μg, approximately 1500 μg to approximately 15000 μg, approximately 1500 μg to approximately 14000 μg 000μg, about 1500μg to about 13000μg, about 1500μg to about 12000μg, about 1500μg to about 11000μg, about 1500μg to about 10000μg, about 1500μg to about 9000μg, about 1500μg to about 8000μg, about 1500μg to about 7000μg, about 1500μg to about 6000μg, about 1500μg to about 5000μg, about 1500μg to about 4500μg, about 1500μg to about 4000μg, about 1500μg to about 3500μg, about 1500μg to about 3000μg, about 1500μg to about 2500μg, about 1500μg to about 2000μg,Approximately 2000 μg to approximately 100000 μg, approximately 2000 μg to approximately 90000 μg, approximately 2000 μg to approximately 80000 μg, approximately 2000 μg to approximately 70000 μg, approximately 2000 μg to approximately 60000 μg, approximately 2000 μg to approximately 50000 μg, approximately 2000 μg to approximately 40000 μg, approximately 2000 μg to approximately 30000 μg, approximately 2000 μg to approximately 29000 μg, approximately 2000 μg to approximately 28000 μg, approximately 2000 μg to approximately 27000 μg, approximately 2000 μg to approximately 26000 μg, approximately 2000 μg to approximately 25000 μg, approximately 2000 μg to approximately 24000 μg, approximately 2000 μg to approximately 23000μg, about 2000μg to about 22000μg, about 2000μg to about 21000μg, about 2000μg to about 20000μg, about 2000μg to about 19000μg, about 2000μg to about 18000μg, about 2000μg to about 17000μg, about 2000μg to about 16000μg, about 2000μg to about 15000μg, about 2000μg to about 14000μg, about 2000μg to about 13000μg, about 2000μg to about 12000μg, about 2000μg to about 11000μg, about 2000μg to about 10000μg, about 2000μg to about 9000μg, about 200 0 μg to about 8000 μg, about 2000 μg to about 7000 μg, about 2000 μg to about 6000 μg, about 2000 μg to about 5000 μg, about 2000 μg to about 4500 μg, about 2000 μg to about 4000 μg, about 2000 μg to about 3500 μg, about 2000 μg to about 3000 μg, about 2000 μg to about 2500 μg, about 2500 μg to about 100000 μg, about 2500 μg to about 90000 μg, about 2500 μg to about 80000 μg, about 2500 μg to about 70000 μg, about 2500 μg to about 60000 μg, about 2500 μg to about 50000 μg, about 2500 μg Approximately 2500 μg to 40000 μg, approximately 2500 μg to 30000 μg, approximately 2500 μg to 29000 μg, approximately 2500 μg to 28000 μg, approximately 2500 μg to 27000 μg, approximately 2500 μg to 26000 μg, approximately 2500 μg to 25000 μg, approximately 2500 μg to 24000 μg, approximately 2500 μg to 23000 μg, approximately 2500 μg to 22000 μg, approximately 2500 μg to 21000 μg, approximately 2500 μg to 20000 μg, approximately 2500 μg to 19000 μg, approximately 2500 μg to 18000 μg, approximately 2500 μg to 17000 μg.Approximately 2500 μg to approximately 16000 μg, approximately 2500 μg to approximately 15000 μg, approximately 2500 μg to approximately 14000 μg, approximately 2500 μg to approximately 13000 μg, approximately 2500 μg to approximately 12000 μg, approximately 2500 μg to approximately 11000 μg, approximately 2500 μg to approximately 10000 μg, approximately 2500 μg to approximately 9000 μg, approximately 2500 μg to approximately 8000 μg, approximately 2500 μg to approximately 7000 μg, approximately 2500 μg to approximately 6000 μg, approximately 2500 μg to approximately 5000 μg, approximately 2500 μg to approximately 4500 μg, approximately 2500 μg to approximately 4000 μg, approximately 2500 μg to approximately 3500 μg. Approximately 2500 μg to approximately 3000 μg, approximately 3000 μg to approximately 100000 μg, approximately 3000 μg to approximately 90000 μg, approximately 3000 μg to approximately 80000 μg, approximately 3000 μg to approximately 70000 μg, approximately 3000 μg to approximately 60000 μg, approximately 3000 μg to approximately 50000 μg, approximately 3000 μg to approximately 40000 μg, approximately 3000 μg to approximately 30000 μg, approximately 3000 μg to approximately 29000 μg, approximately 3000 μg to approximately 28000 μg, approximately 3000 μg to approximately 27000 μg, approximately 3000 μg to approximately 26000 μg, approximately 3000 μg to approximately 25000 μg, approximately 3000 μg to approximately 24000 μg, about 3000 μg to about 23000 μg, about 3000 μg to about 22000 μg, about 3000 μg to about 21000 μg, about 3000 μg to about 20000 μg, about 3000 μg to about 19000 μg, about 3000 μg to about 18000 μg, about 3000 μg to about 17000 μg, about 3000 μg to about 16000 μg, about 3000 μg to about 15000 μg, about 3000 μg to about 14000 μg, about 3000 μg to about 13000 μg, about 3000 μg to about 12000 μg, about 3000 μg to about 11000 μg, about 3000 μg to about 10000 μg, about 3000μg to about 9000μg, about 3000μg to about 8000μg, about 3000μg to about 7000μg, about 3000μg to about 6000μg, about 3000μg to about 5000μg, about 3000μg to about 4500μg, about 3000μg to about 4000μg, about 3500μg to about 100000μg, about 3500μg to about 90000μg, about 3500μg to about 80000μg, about 3500μg to about 70000μg, about 3500μg to about 60000μg, about 3500μg to about 50000μg, about 3500μg to about 40000μg, about 3500μg to about 30000μgApproximately 3500 μg to approximately 29000 μg, approximately 3500 μg to approximately 28000 μg, approximately 3500 μg to approximately 27000 μg, approximately 3500 μg to approximately 26000 μg, approximately 3500 μg to approximately 25000 μg, approximately 3500 μg to approximately 24000 μg, approximately 3500 μg to approximately 23000 μg, approximately 3500 μg to approximately 22000 μg, approximately 3500 μg to approximately 21000 μg, approximately 3500 μg to approximately 20000 μg, approximately 3500 μg to approximately 19000 μg, approximately 3500 μg to approximately 18000 μg, Approximately 3500 μg to approximately 17000 μg, approximately 3500 μg to approximately 16000 μg, approximately 3500 μg to approximately 15000 μg, approximately 3500 μg to approximately 14000 μg, approximately 3500 μg to approximately 13000 μg, approximately 3500 μg to approximately 12000 μg, approximately 3500 μg to approximately 11000 μg, approximately 3500 μg to approximately 10000 μg, approximately 3500 μg to approximately 9000 μg, approximately 3500 μg to approximately 8000 μg, approximately 3500 μg to approximately 7000 μg, approximately 3500 μg to approximately 6000 μg (about) 3500μg to about 5000μg, about 3500μg to about 4500μg, about 3500μg to about 4000μg, about 4000μg to about 100000μg, about 4000μg to about 90000μg, about 4000μg to about 80000μg, about 4000μg to about 70000μg, about 4000μg to about 60000μg, about 4000μg to about 50000μg μg, about 4000μg to about 40000μg, about 4000μg to about 30000μg, about 4000μg to about 29000μg, about 4000μg to about 28000μg, about 4000μg to about 27000μg, about 4000μg to about 26000μg, about 4000μg to about 25000μg, about 4000μg to about 24000μg, about 4000μg to Approximately 23,000 μg, approximately 4,000 μg to approximately 22,000 μg, approximately 4,000 μg to approximately 21,000 μg, approximately 4,000 μg to approximately 20,000 μg, approximately 4,000 μg to approximately 19,000 μg, approximately 4,000 μg to approximately 18,000 μg, approximately 4,000 μg to approximately 17,000 μg, approximately 4,000 μg to approximately 16,000 μg, approximately 4,000 μg to approximately 15,000 μg, approximately 4 000μg to about 14000μg, about 4000μg to about 13000μg, about 4000μg to about 12000μg, about 4000μg to about 11000μg, about 4000μg to about 10000μg, about 4000μg to about 9000μg, about 4000μg to about 8000μg, about 4000μg to about 7000μg, about 4000μg to about 6000μg,Approximately 4000 μg to approximately 5000 μg, approximately 4000 μg to approximately 4500 μg, approximately 4500 μg to approximately 100000 μg, approximately 4500 μg to approximately 90000 μg, approximately 4500 μg to approximately 80000 μg, approximately 4500 μg to approximately 70000 μg, approximately 4500 μg to approximately 60000 μg, approximately 4500 μg to approximately 50000 μg, approximately 4500 μg to approximately 40000 μg, approximately 4500 μg to approximately 30000 μg, approximately 4500 μg to approximately 29000 μg, approximately 4500 μg to approximately 28000 μg, approximately 4500 μg to approximately 27000 μg, approximately 4500 μg to approximately 26000 μg, approximately 4500 μg to approximately 250 00 μg, about 4500 μg to about 24000 μg, about 4500 μg to about 23000 μg, about 4500 μg to about 22000 μg, about 4500 μg to about 21000 μg, about 4500 μg to about 20000 μg, about 4500 μg to about 19000 μg, about 4500 μg to about 18000 μg, about 4500 μg to about 17000 μg, about 4500 μg to about 16000 μg, about 4500 μg to about 15000 μg, about 4500 μg to about 14000 μg, about 4500 μg to about 13000 μg, about 4500 μg to about 12000 μg, about 4500 μg to about 11000 μg, about 4500 μg Approximately 10,000 μg, approximately 4,500 μg to approximately 9,000 μg, approximately 4,500 μg to approximately 8,000 μg, approximately 4,500 μg to approximately 7,000 μg, approximately 4,500 μg to approximately 6,000 μg, approximately 4,500 μg to approximately 5,000 μg, approximately 5,000 μg to approximately 100,000 μg, approximately 5,000 μg to approximately 90,000 μg, approximately 5,000 μg to approximately 80,000 μg, approximately 5,000 μg to approximately 70,000 μg, approximately 5,000 μg to approximately 60,000 μg, approximately 5,000 μg to approximately 50,000 μg, approximately 5,000 μg to approximately 40,000 μg, approximately 5,000 μg to approximately 30,000 μg, approximately 5,000 μg to approximately 29,000 μg, approximately 5,000 μg Approximately 5000 μg to 28000 μg, approximately 5000 μg to 27000 μg, approximately 5000 μg to 26000 μg, approximately 5000 μg to 25000 μg, approximately 5000 μg to 24000 μg, approximately 5000 μg to 23000 μg, approximately 5000 μg to 22000 μg, approximately 5000 μg to 21000 μg, approximately 5000 μg to 20000 μg, approximately 5000 μg to 19000 μg, approximately 5000 μg to 18000 μg, approximately 5000 μg to 17000 μg, approximately 5000 μg to 16000 μg, approximately 5000 μg to 15000 μg, approximately 5000 μg to 14000 μg.Approximately 5000 μg to approximately 13000 μg, approximately 5000 μg to approximately 12000 μg, approximately 5000 μg to approximately 11000 μg, approximately 5000 μg to approximately 10000 μg, approximately 5000 μg to approximately 9000 μg, approximately 5000 μg to approximately 8000 μg, approximately 5000 μg to approximately 7000 μg, approximately 5000 μg to approximately 6000 μg, approximately 6000 μg to approximately 100000 μg, approximately 6000 μg to approximately 90000 μg, approximately 6000 μg to approximately 80000 μg, approximately 6000 μg to approximately 70000 μg, approximately 6000 μg to approximately 60000 μg, approximately 6000 μg to approximately 50000 μg, approximately 6000 μg to approximately 40000 μg μg, about 6000μg to about 30000μg, about 6000μg to about 29000μg, about 6000μg to about 28000μg, about 6000μg to about 27000μg, about 6000μg to about 26000μg, about 6000μg to about 25000μg, about 6000μg to about 24000μg, about 6000μg to about 23000μg, about 6000μg to about 22000μg, about 6000μg to about 21000μg, about 6000μg to about 20000μg, about 6000μg to about 19000μg, about 6000μg to about 18000μg, about 6000μg to about 17000μg, about 6000μg to about 16000μg, about 6000μg to about 15000μg, about 6000μg to about 14000μg, about 6000μg to about 13000μg, about 6000μg to about 12000μg, about 6000μg to about 11000μg, about 6000μg to about 10000μg, about 6000μg to about 9000μg, about 6000μg to about 8000μg, about 6000μg to about 7000μg, about 7000μg to about 100000μg, about 7000μg to about 90000μg, about 7000μg to about 80000μg, about 7000μg to about 70000μg, about 7000μg to about 60000μg, about 7000μg to about 60000μg, about 7000μg to about 70000μg. Approximately 7000 μg to 50000 μg, approximately 7000 μg to 40000 μg, approximately 7000 μg to 30000 μg, approximately 7000 μg to 29000 μg, approximately 7000 μg to 28000 μg, approximately 7000 μg to 27000 μg, approximately 7000 μg to 26000 μg, approximately 7000 μg to 25000 μg, approximately 7000 μg to 24000 μg, approximately 7000 μg to 23000 μg, approximately 7000 μg to 22000 μg, approximately 7000 μg to 21000 μg, approximately 7000 μg to 20000 μg, approximately 7000 μg to 19000 μg, approximately 7000 μg to 18000 μg.Approximately 7000 μg to approximately 17000 μg, approximately 7000 μg to approximately 16000 μg, approximately 7000 μg to approximately 15000 μg, approximately 7000 μg to approximately 14000 μg, approximately 7000 μg to approximately 13000 μg, approximately 7000 μg to approximately 12000 μg, approximately 7000 μg to approximately 11000 μg, approximately 7000 μg to approximately 10000 μg, approximately 7000 μg to approximately 9000 μg, approximately 7000 μg to approximately 8000 μg, approximately 8000 μg to approximately 100000 μg, approximately 8000 μg to approximately 90000 μg, approximately 8000 μg to approximately 80000 μg, approximately 8000 μg to approximately 70000 μg, approximately 8000 μg to approximately 60,000 μg, about 8,000 μg to about 50,000 μg, about 8,000 μg to about 40,000 μg, about 8,000 μg to about 30,000 μg, about 8,000 μg to about 29,000 μg, about 8,000 μg to about 28,000 μg, about 8,000 μg to about 27,000 μg, about 8,000 μg to about 26,000 μg, about 8,000 μg to about 25,000 μg, about 8,000 μg to about 24,000 μg, about 8,000 μg to about 23,000 μg, about 8,000 μg to about 22,000 μg, about 8,000 μg to about 21,000 μg, about 8,000 μg to about 20,000 μg, about 8,000 μg to about 19,000 μg. Approximately 8000 μg to approximately 18000 μg, approximately 8000 μg to approximately 17000 μg, approximately 8000 μg to approximately 16000 μg, approximately 8000 μg to approximately 15000 μg, approximately 8000 μg to approximately 14000 μg, approximately 8000 μg to approximately 13000 μg, approximately 8000 μg to approximately 12000 μg, approximately 8000 μg to approximately 11000 μg, approximately 8000 μg to approximately 10000 μg, approximately 8000 μg to approximately 9000 μg, approximately 9000 μg to approximately 100000 μg, approximately 9000 μg to approximately 90000 μg, approximately 9000 μg to approximately 80000 μg, approximately 9000 μg to approximately 70000 μg, approximately 9000 μg to Approximately 60,000 μg, approximately 9,000 μg to approximately 50,000 μg, approximately 9,000 μg to approximately 40,000 μg, approximately 9,000 μg to approximately 30,000 μg, approximately 9,000 μg to approximately 29,000 μg, approximately 9,000 μg to approximately 28,000 μg, approximately 9,000 μg to approximately 27,000 μg, approximately 9,000 μg to approximately 26,000 μg, approximately 9,000 μg to approximately 25,000 μg, approximately 9,000 μg to approximately 24,000 μg, approximately 9,000 μg to approximately 23,000 μg, approximately 9,000 μg to approximately 22,000 μg, approximately 9,000 μg to approximately 21,000 μg, approximately 9,000 μg to approximately 20,000 μg, approximately 9,000 μg to approximately 19,000 μg.Approximately 9000 μg to approximately 18000 μg, approximately 9000 μg to approximately 17000 μg, approximately 9000 μg to approximately 16000 μg, approximately 9000 μg to approximately 15000 μg, approximately 9000 μg to approximately 14000 μg, approximately 9000 μg to approximately 13000 μg, approximately 9000 μg to approximately 12000 μg, approximately 9000 μg to approximately 11000 μg, approximately 9000 μg to approximately 10000 μg, approximately 10000 μg to approximately 100000 μg, approximately 10000 μg to approximately 90000 μg, approximately 10000 μg to approximately 80000 μg, approximately 10000 μg to approximately 70000 μg, approximately 10000 μg to approximately 60000 μg, approximately 10000 μg to approximately 60000 μg, approximately 10000 μg to approximately 18000 μg, approximately 18000 μg to approximately 17000 μg, approximately 17 ... μg to about 50,000 μg, about 10,000 μg to about 40,000 μg, about 10,000 μg to about 30,000 μg, about 10,000 μg to about 29,000 μg, about 10,000 μg to about 28,000 μg, about 10,000 μg to about 27,000 μg, about 10,000 μg to about 26,000 μg, about 10,000 μg to about 25,000 μg, about 10,000 μg to about 24,000 μg, about 10,000 μg to about 23,000 μg, about 10,000 μg to about 22,000 μg, about 10,000 μg to about 21,000 μg, about 10,000 μg to about 20,000 μg, about 10,000 μg to about 19,000 μg, about 100 0.00 μg to about 18000 μg, about 10000 μg to about 17000 μg, about 10000 μg to about 16000 μg, about 10000 μg to about 15000 μg, about 10000 μg to about 14000 μg, about 10000 μg to about 13000 μg, about 10000 μg to about 12000 μg, about 10000 μg to about 11000 μg, about 11000 μg to about 100000 μg, about 11000 μg to about 90000 μg, about 11000 μg to about 80000 μg, about 11000 μg to about 70000 μg, about 11000 μg to about 60000 μg, about 11000 μg to about 50000 μg, about 11,000 μg to about 40,000 μg, about 11,000 μg to about 30,000 μg, about 11,000 μg to about 29,000 μg, about 11,000 μg to about 28,000 μg, about 11,000 μg to about 27,000 μg, about 11,000 μg to about 26,000 μg, about 11,000 μg to about 25,000 μg, about 11,000 μg to about 24,000 μg, about 11,000 μg to about 23,000 μg, about 11,000 μg to about 22,000 μg, about 11,000 μg to about 21,000 μg, about 11,000 μg to about 20,000 μg, about 11,000 μg to about 19,000 μg, about 11,000 μg to about 18,000 μg.Approximately 11,000 μg to approximately 17,000 μg, approximately 11,000 μg to approximately 16,000 μg, approximately 11,000 μg to approximately 15,000 μg, approximately 11,000 μg to approximately 14,000 μg, approximately 11,000 μg to approximately 13,000 μg, approximately 11,000 μg to approximately 12,000 μg, approximately 12,000 μg to approximately 100,000 μg, approximately 12,000 μg to approximately 90,000 μg, approximately 12,000 μg to approximately 80,000 μg, approximately 12,000 μg to approximately 70,000 μg, approximately 12,000 μg to approximately 60,000 μg, approximately 12,000 μg to approximately 50,000 μg, approximately 12,000 μg to approximately 40,000 μg, approximately 12,000 μg to approximately 300 μg 0.00 μg, about 12,000 μg to about 29,000 μg, about 12,000 μg to about 28,000 μg, about 12,000 μg to about 27,000 μg, about 12,000 μg to about 26,000 μg, about 12,000 μg to about 25,000 μg, about 12,000 μg to about 24,000 μg, about 12,000 μg to about 23,000 μg, about 12,000 μg to about 22,000 μg, about 12,000 μg to about 21,000 μg, about 12,000 μg to about 20,000 μg, about 12,000 μg to about 19,000 μg, about 12,000 μg to about 18,000 μg, about 12,000 μg to about 17,000 μg, about 12,000 μg to Approximately 16,000 μg, approximately 12,000 μg to approximately 15,000 μg, approximately 12,000 μg to approximately 14,000 μg, approximately 12,000 μg to approximately 13,000 μg, approximately 13,000 μg to approximately 100,000 μg, approximately 13,000 μg to approximately 90,000 μg, approximately 13,000 μg to approximately 80,000 μg, approximately 13,000 μg to approximately 70,000 μg, approximately 13,000 μg to approximately 60,000 μg, approximately 13,000 μg to approximately 50,000 μg, approximately 13,000 μg to approximately 40,000 μg, approximately 13,000 μg to approximately 30,000 μg, approximately 13,000 μg to approximately 29,000 μg, approximately 13,000 μg to approximately 28,000 μg, approximately 130 0.00 μg to about 27000 μg, about 13000 μg to about 26000 μg, about 13000 μg to about 25000 μg, about 13000 μg to about 24000 μg, about 13000 μg to about 23000 μg, about 13000 μg to about 22000 μg, about 13000 μg to about 21000 μg, about 13000 μg to about 20000 μg, about 13000 μg to about 19000 μg, about 13000 μg to about 18000 μg, about 13000 μg to about 17000 μg, about 13000 μg to about 16000 μg, about 13000 μg to about 15000 μg, about 13000 μg to about 14000 μg,Approximately 14,000 μg to approximately 100,000 μg, approximately 14,000 μg to approximately 90,000 μg, approximately 14,000 μg to approximately 80,000 μg, approximately 14,000 μg to approximately 70,000 μg, approximately 14,000 μg to approximately 60,000 μg, approximately 14,000 μg to approximately 50,000 μg, approximately 14,000 μg to approximately 40,000 μg, approximately 14,000 μg to approximately 30,000 μg, approximately 14,000 μg to approximately 29,000 μg, approximately 14,000 μg to approximately 28,000 μg, approximately 14,000 μg to approximately 27,000 μg, approximately 14,000 μg to approximately 26,000 μg, approximately 14,000 μg to approximately 25,000 μg, approximately 14,000 μg to approximately 240 μg 0.00 μg, about 14,000 μg to about 23,000 μg, about 14,000 μg to about 22,000 μg, about 14,000 μg to about 21,000 μg, about 14,000 μg to about 20,000 μg, about 14,000 μg to about 19,000 μg, about 14,000 μg to about 18,000 μg, about 14,000 μg to about 17,000 μg, about 14,000 μg to about 16,000 μg, about 14,000 μg to about 15,000 μg, about 15,000 μg to about 100,000 μg, about 15,000 μg to about 90,000 μg, about 15,000 μg to about 80,000 μg, about 15,000 μg to about 70,000 μg, about 15,000 μg Approximately 60,000 μg, approximately 15,000 μg to approximately 50,000 μg, approximately 15,000 μg to approximately 40,000 μg, approximately 15,000 μg to approximately 30,000 μg, approximately 15,000 μg to approximately 29,000 μg, approximately 15,000 μg to approximately 28,000 μg, approximately 15,000 μg to approximately 27,000 μg, approximately 15,000 μg to approximately 26,000 μg, approximately 15,000 μg to approximately 25,000 μg, approximately 15,000 μg to approximately 24,000 μg, approximately 15,000 μg to approximately 23,000 μg, approximately 15,000 μg to approximately 22,000 μg, approximately 15,000 μg to approximately 21,000 μg, approximately 15,000 μg to approximately 20,000 μg, approximately 150 0.00 μg to about 19000 μg, about 15000 μg to about 18000 μg, about 15000 μg to about 17000 μg, about 15000 μg to about 16000 μg, about 16000 μg to about 100000 μg, about 16000 μg to about 90000 μg, about 16000 μg to about 80000 μg, about 16000 μg to about 70000 μg, about 16000 μg to about 60000 μg, about 16000 μg to about 50000 μg, about 16000 μg to about 40000 μg, about 16000 μg to about 30000 μg, about 16000 μg to about 29000 μg, about 16000 μg to about 28000 μg,Approximately 16,000 μg to approximately 27,000 μg, approximately 16,000 μg to approximately 26,000 μg, approximately 16,000 μg to approximately 25,000 μg, approximately 16,000 μg to approximately 24,000 μg, approximately 16,000 μg to approximately 23,000 μg, approximately 16,000 μg to approximately 22,000 μg, approximately 16,000 μg to approximately 21,000 μg, approximately 16,000 μg to approximately 20,000 μg, approximately 16,000 μg to approximately 19,000 μg, approximately 16,000 μg to approximately 18,000 μg, approximately 16,000 μg to approximately 17,000 μg, approximately 17,000 μg to approximately 100,000 μg, approximately 17,000 μg to approximately 90,000 μg, approximately 17,000 μg to approximately 800 μg 0.00 μg, about 17,000 μg to about 70,000 μg, about 17,000 μg to about 60,000 μg, about 17,000 μg to about 50,000 μg, about 17,000 μg to about 40,000 μg, about 17,000 μg to about 30,000 μg, about 17,000 μg to about 29,000 μg, about 17,000 μg to about 28,000 μg, about 17,000 μg to about 27,000 μg, about 17,000 μg to about 26,000 μg, about 17,000 μg to about 25,000 μg, about 17,000 μg to about 24,000 μg, about 17,000 μg to about 23,000 μg, about 17,000 μg to about 22,000 μg, about 17,000 μg to Approximately 21,000 μg, approximately 17,000 μg to approximately 20,000 μg, approximately 17,000 μg to approximately 19,000 μg, approximately 17,000 μg to approximately 18,000 μg, approximately 18,000 μg to approximately 100,000 μg, approximately 18,000 μg to approximately 90,000 μg, approximately 18,000 μg to approximately 80,000 μg, approximately 18,000 μg to approximately 70,000 μg, approximately 18,000 μg to approximately 60,000 μg, approximately 18,000 μg to approximately 50,000 μg, approximately 18,000 μg to approximately 40,000 μg, approximately 18,000 μg to approximately 30,000 μg, approximately 18,000 μg to approximately 29,000 μg, approximately 18,000 μg to approximately 28,000 μg, approximately 180 0.00 μg to about 27000 μg, about 18000 μg to about 26000 μg, about 18000 μg to about 25000 μg, about 18000 μg to about 24000 μg, about 18000 μg to about 23000 μg, about 18000 μg to about 22000 μg, about 18000 μg to about 21000 μg, about 18000 μg to about 20000 μg, about 18000 μg to about 19000 μg, about 19000 μg to about 100000 μg, about 19000 μg to about 90000 μg, about 19000 μg to about 80000 μg, about 19000 μg to about 70000 μg, about 19000 μg to about 60000 μg,Approximately 19,000 μg to approximately 50,000 μg, approximately 19,000 μg to approximately 40,000 μg, approximately 19,000 μg to approximately 30,000 μg, approximately 19,000 μg to approximately 29,000 μg, approximately 19,000 μg to approximately 28,000 μg, approximately 19,000 μg to approximately 27,000 μg, approximately 19,000 μg to approximately 26,000 μg, approximately 19,000 μg to approximately 25,000 μg, approximately 19,000 μg to approximately 24,000 μg, approximately 19,000 μg to approximately 23,000 μg, approximately 19,000 μg to approximately 22,000 μg, approximately 19,000 μg to approximately 21,000 μg, approximately 19,000 μg to approximately 20,000 μg, approximately 20,000 μg to approximately 1,000 μg 0.00 μg, about 20,000 μg to about 90,000 μg, about 20,000 μg to about 80,000 μg, about 20,000 μg to about 70,000 μg, about 20,000 μg to about 60,000 μg, about 20,000 μg to about 50,000 μg, about 20,000 μg to about 40,000 μg, about 20,000 μg to about 30,000 μg, about 20,000 μg to about 29,000 μg, about 20,000 μg to about 28,000 μg, about 20,000 μg to about 27,000 μg, about 20,000 μg to about 26,000 μg, about 20,000 μg to about 25,000 μg, about 20,000 μg to about 24,000 μg, about 20,000 μg to Approximately 23,000 μg, approximately 20,000 μg to approximately 22,000 μg, approximately 20,000 μg to approximately 21,000 μg, approximately 21,000 μg to approximately 100,000 μg, approximately 21,000 μg to approximately 90,000 μg, approximately 21,000 μg to approximately 80,000 μg, approximately 21,000 μg to approximately 70,000 μg, approximately 21,000 μg to approximately 60,000 μg, approximately 21,000 μg to approximately 50,000 μg, approximately 21,000 μg to approximately 40,000 μg, approximately 21,000 μg to approximately 30,000 μg, approximately 21,000 μg to approximately 29,000 μg, approximately 21,000 μg to approximately 28,000 μg, approximately 21,000 μg to approximately 27,000 μg, approximately 210 0.00 μg to about 26000 μg, about 21000 μg to about 25000 μg, about 21000 μg to about 24000 μg, about 21000 μg to about 23000 μg, about 21000 μg to about 22000 μg, about 22000 μg to about 100000 μg, about 22000 μg to about 90000 μg, about 22000 μg to about 80000 μg, about 22000 μg to about 70000 μg, about 22000 μg to about 60000 μg, about 22000 μg to about 50000 μg, about 22000 μg to about 40000 μg, about 22000 μg to about 30000 μg, about 22000 μg to about 29000 μg,Approximately 22,000 μg to approximately 28,000 μg, approximately 22,000 μg to approximately 27,000 μg, approximately 22,000 μg to approximately 26,000 μg, approximately 22,000 μg to approximately 25,000 μg, approximately 22,000 μg to approximately 24,000 μg, approximately 22,000 μg to approximately 23,000 μg, approximately 23,000 μg to approximately 100,000 μg, approximately 23,000 μg to approximately 90,000 μg, approximately 23,000 μg to approximately 80,000 μg, approximately 23,000 μg to approximately 70,000 μg, approximately 23,000 μg to approximately 60,000 μg, approximately 23,000 μg to approximately 50,000 μg, approximately 23,000 μg to approximately 40,000 μg, approximately 23,000 μg to approximately 300 μg 0.00 μg, about 23,000 μg to about 29,000 μg, about 23,000 μg to about 28,000 μg, about 23,000 μg to about 27,000 μg, about 23,000 μg to about 26,000 μg, about 23,000 μg to about 25,000 μg, about 23,000 μg to about 24,000 μg, about 24,000 μg to about 100,000 μg, about 24,000 μg to about 90,000 μg, about 24,000 μg to about 80,000 μg, about 24,000 μg to about 70,000 μg, about 24,000 μg to about 60,000 μg, about 24,000 μg to about 50,000 μg, about 24,000 μg to about 40,000 μg, about 24,000 μg to Approximately 30,000 μg, approximately 24,000 μg to approximately 29,000 μg, approximately 24,000 μg to approximately 28,000 μg, approximately 24,000 μg to approximately 27,000 μg, approximately 24,000 μg to approximately 26,000 μg, approximately 24,000 μg to approximately 25,000 μg, approximately 25,000 μg to approximately 100,000 μg, approximately 25,000 μg to approximately 90,000 μg, approximately 25,000 μg to approximately 80,000 μg, approximately 25,000 μg to approximately 70,000 μg, approximately 25,000 μg to approximately 60,000 μg, approximately 25,000 μg to approximately 50,000 μg, approximately 25,000 μg to approximately 40,000 μg, approximately 25,000 μg to approximately 30,000 μg, approximately 250 0.00 μg to about 29,000 μg, about 25,000 μg to about 28,000 μg, about 25,000 μg to about 27,000 μg, about 25,000 μg to about 26,000 μg, about 26,000 μg to about 100,000 μg, about 26,000 μg to about 90,000 μg, about 26,000 μg to about 80,000 μg, about 26,000 μg to about 70,000 μg, about 26,000 μg to about 60,000 μg, about 26,000 μg to about 50,000 μg, about 26,000 μg to about 40,000 μg, about 26,000 μg to about 30,000 μg, about 26,000 μg to about 29,000 μg, about 26,000 μg to about 28,000 μg,Approximately 26,000 μg to approximately 27,000 μg, approximately 27,000 μg to approximately 100,000 μg, approximately 27,000 μg to approximately 90,000 μg, approximately 27,000 μg to approximately 80,000 μg, approximately 27,000 μg to approximately 70,000 μg, approximately 27,000 μg to approximately 60,000 μg, approximately 27,000 μg to approximately 50,000 μg, approximately 27,000 μg to approximately 40,000 μg, approximately 27,000 μg to approximately 30,000 μg, approximately 27,000 μg to approximately 29,000 μg, approximately 27,000 μg to approximately 28,000 μg, approximately 28,000 μg to approximately 100,000 μg, approximately 28,000 μg to approximately 90,000 μg, approximately 28,000 μg to Approximately 80,000 μg, approximately 28,000 μg to approximately 70,000 μg, approximately 28,000 μg to approximately 60,000 μg, approximately 28,000 μg to approximately 50,000 μg, approximately 28,000 μg to approximately 40,000 μg, approximately 28,000 μg to approximately 30,000 μg, approximately 28,000 μg to approximately 29,000 μg, approximately 29,000 μg to approximately 100,000 μg, approximately 29,000 μg to approximately 90,000 μg, approximately 29,000 μg to approximately 80,000 μg, approximately 29,000 μg to approximately 70,000 μg, approximately 29,000 μg to approximately 60,000 μg, approximately 29,000 μg to approximately 50,000 μg, approximately 29,000 μg to approximately 40,000 μg, approximately 29,000 μg to about 30,000 μg, about 30,000 μg to about 100,000 μg, about 30,000 μg to about 90,000 μg, about 30,000 μg to about 80,000 μg, about 30,000 μg to about 70,000 μg, about 30,000 μg to about 60,000 μg, about 30,000 μg to about 50,000 μg, about 30,000 μg to about 40,000 μg, about 40,000 μg to about 100,000 μg, about 40,000 μg to about 90,000 μg, about 40,000 μg to about 80,000 μg, about 40,000 μg to about 70,000 μg, about 40,000 μg to about 60,000 μg, about 40,000 μg to about 50,000 μg, about 50,000 μg to about 100,000 μg, about 50,000 μg to about 90,000 μg, about 50,000 μg to about 80,000 μg, about 50,000 μg to about 70,000 μg, about 50,000 μg to about 60,000 μg, about 60,000 μg to about 100,000 μg, about 60,000 μg to about 90,000 μg, about 60,000 μg to about 80,000 μg, about 60,000 μg to about 70,000 μg, about 70,000 μg to about 100,000 μg, about 70,000 μg to about 90,000 μg, about 70,000 μg to about 80,000 μg, about 80,000 μg to about 100,000 μgApproximately 80,000 μg to approximately 90,000 μg and approximately 90,000 μg to approximately 100,000 μg.

[0195] In some embodiments, the dosage of the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) may be from about 1.0 μg to about 10,000 μg, from about 1.0 μg to about 5,000 μg, from about 1.0 μg to about 2,500 μg, from about 1.0 μg to about 1,000 μg, from about 1.0 μg to about 500 μg, from about 1.0 μg to about 250 μg, from about 1.0 μg to about 100 μg, from about 1.0 μg to about 75 μg, from about 1.0 μg to about 50 μg, from about 1.0 μg to about 25 μg, from about 1.0 μg to about 10 μg, from about 10 μg to about 10,000 μg, from about 10 μg to about 5,000 μg, from about 10 μg to about 2,500 μg, from about 10 μg to about 10 ...10000 μg, from about 10 μg to about 2500 μg, from about 10 μg to about 10000 μg, from about 10 μg to about 10000 μg, from about 10 μg to about 2500 μg, from about 10 μg to about 10000 μg, from about 10 μg to about 10000 μg, from about 10 μg to about 10000 μg, from about 10 μg to about 1000 000μg, about 10μg to about 500μg, about 10μg to about 250μg, about 10μg to about 100μg, about 10μg to about 75μg, about 10μg to about 50μg, about 50μg to about 10000μg, about 50μg to about 5000μg, about 50μg to about 2500μg, about 50μg to about 2000μg, about 50μg to about 1000μg, about 50μg to about 750μg, about 50μg to about 5000μg, about 50μg to about 2500μg, about 50μg to about 2000μg, about 50μg to about 1000μg, about 100μg to about 10000μg, about 100μg to about 5000μg, about 100μg to about 2500μg μg, about 100μg to about 2000μg, about 100μg to about 1500μg, about 100μg to about 1250μg, about 100μg to about 1000μg, about 100μg to about 750μg, about 100μg to about 500μg, about 100μg to about 250μg, about 100μg to about 200μg, about 250μg to about 10000μg, about 250μg to about 5000μg, about 250μg to about 2500μg, about 250μg to about 2000μg, about 250μg to about 1500μg, about 250μg to about 1250μg, about 250μg to about 1000μg, about 250μg to about 750μg, about 250μg to about 5 0.00 μg, about 500 μg to about 10000 μg, about 500 μg to about 7000 μg, about 500 μg to about 5000 μg, about 500 μg to about 3000 μg, about 500 μg to about 2500 μg, about 500 μg to about 2000 μg, about 500 μg to about 1500 μg, about 500 μg to about 1250 μg, about 500 μg to about 1000 μg, about 500 μg to about 750 μg, about 750 μg to about 20000 μg, about 750 μg to about 15000 μg, about 750 μg to about 10000 μg, about 750 μg to about 7000 μg, about 750 μg to about 5000 μg, about 750 μg to about 3000 μg,Approximately 750 μg to approximately 2500 μg, approximately 750 μg to approximately 2000 μg, approximately 750 μg to approximately 1500 μg, approximately 750 μg to approximately 1250 μg, approximately 750 μg to approximately 1000 μg, approximately 1000 μg to approximately 25000 μg, approximately 1000 μg to approximately 20000 μg, approximately 1000 μg to approximately 15000 μg, approximately 1000 μg to approximately 10000 μg, approximately 1000 μg to approximately 7000 μg, approximately 1000 μg to approximately 5000 μg, approximately 1000 μg to approximately 3000 μg, approximately 1000 μg to approximately 2500 μg, approximately 1000 μg to approximately 2000 μg, approximately 1000 μg to approximately 1500 μg, approximately 1500 μg About 25,000 μg, about 1,500 μg to about 20,000 μg, about 1,500 μg to about 15,000 μg, about 1,500 μg to about 10,000 μg, about 1,500 μg to about 7,000 μg, about 1,500 μg to about 5,000 μg, about 1,500 μg to about 3,000 μg, about 1,500 μg to about 2,500 μg, about 1,500 μg to about 2,000 μg, about 2,000 μg to about 30,000 μg, about 2,000 μg to about 25,000 μg, about 2,000 μg to about 20,000 μg, about 2,000 μg to about 15,000 μg, about 2,000 μg to about 10,000 μg, about 2,000 μg to about 7,000 μg, about 2,000 μg μg to about 5000μg, about 2000μg to about 4000μg, about 2000μg to about 3000μg, about 2000μg to about 2500μg, about 2500μg to about 30000μg, about 2500μg to about 25000μg, about 2500μg to about 20000μg, about 2500μg to about 15000μg, about 2500μg to about 12000μg, about 2500μg to about 10000μg, about 2500μg to about 7000μg, about 2500μg to about 5000μg, about 2500μg to about 4000μg, about 2500μg to about 3000μg, about 3000μg to about 50000μg, about 300μg 0 μg to about 25000 μg, about 3000 μg to about 20000 μg, about 3000 μg to about 15000 μg, about 3000 μg to about 12000 μg, about 3000 μg to about 10000 μg, about 3000 μg to about 7000 μg, about 3000 μg to about 6000 μg, about 3000 μg to about 5000 μg, about 3000 μg to about 4000 μg, about 4000 μg to about 50000 μg, about 4000 μg to about 30000 μg, about 4000 μg to about 25000 μg, about 4000 μg to about 20000 μg, about 4000 μg to about 15000 μg, about 4000 μg to about 12000 μg,Approximately 4000 μg to approximately 10000 μg, approximately 4000 μg to approximately 8000 μg, approximately 4000 μg to approximately 6000 μg, approximately 4000 μg to approximately 5000 μg, approximately 5000 μg to approximately 50000 μg, approximately 5000 μg to approximately 40000 μg, approximately 5000 μg to approximately 30000 μg, approximately 5000 μg to approximately 25000 μg, approximately 5000 μg to approximately 20000 μg, approximately 5000 μg to approximately 15000 μg, approximately 5000 μg to approximately 12000 μg, approximately 5000 μg to approximately 10000 μg, approximately 5000 μg to approximately 8000 μg, approximately 5000 μg to approximately 7000 μg, approximately 5000 μg to approximately 6000 μg μg, about 6000μg to about 50000μg, about 6000μg to about 30000μg, about 6000μg to about 25000μg, about 6000μg to about 20000μg, about 6000μg to about 17000μg, about 6000μg to about 15000μg, about 6000μg to about 12000μg, about 6000μg to about 10000μg, about 6000μg to about 9000μg, about 6000μg to about 8000μg, about 6000μg to about 7000μg, about 7000μg to about 70000μg, about 7000μg to about 50000μg, about 7000μg to about 40000μg, about 7000μg to about 30,000 μg, about 7,000 μg to about 25,000 μg, about 7,000 μg to about 20,000 μg, about 7,000 μg to about 17,000 μg, about 7,000 μg to about 15,000 μg, about 7,000 μg to about 12,000 μg, about 7,000 μg to about 10,000 μg, about 7,000 μg to about 8,000 μg, about 8,000 μg to about 80,000 μg, about 8,000 μg to about 60,000 μg, about 8,000 μg to about 50,000 μg, about 8,000 μg to about 30,000 μg, about 8,000 μg to about 25,000 μg, about 8,000 μg to about 20,000 μg, about 8,000 μg to about 17,000 μg, about 8 000μg to about 15000μg, about 8000μg to about 12000μg, about 8000μg to about 10000μg, about 10000μg to about 100000μg, about 10000μg to about 80000μg, about 10000μg to about 70000μg, about 10000μg to about 60000μg, about 10000μg to about 50000μg, about 10000μg to about 40000μg, about 10000μg to about 30000μg, about 10000μg to about 25000μg, about 10000μg to about 20000μg, about 10000μg to about 18000μg, about 10000μg to about 16000μg,Approximately 10,000 μg to approximately 15,000 μg, approximately 10,000 μg to approximately 12,000 μg, approximately 12,000 μg to approximately 100,000 μg, approximately 12,000 μg to approximately 90,000 μg, approximately 12,000 μg to approximately 80,000 μg, approximately 12,000 μg to approximately 70,000 μg, approximately 12,000 μg to approximately 60,000 μg, approximately 12,000 μg to approximately 50,000 μg, approximately 12,000 μg to approximately 40,000 μg, approximately 12,000 μg to approximately 30,000 μg, approximately 12,000 μg to approximately 25,000 μg, approximately 12,000 μg to approximately 20,000 μg, approximately 12,000 μg to approximately 18,000 μg, approximately 12,000 μg to approximately 1600 μg 0 μg, about 12,000 μg to about 15,000 μg, about 12,000 μg to about 14,000 μg, about 15,000 μg to about 100,000 μg, about 15,000 μg to about 90,000 μg, about 15,000 μg to about 80,000 μg, about 15,000 μg to about 70,000 μg, about 15,000 μg to about 60,000 μg, about 15,000 μg to about 50,000 μg, about 15,000 μg to about 40,000 μg, about 15,000 μg to about 30,000 μg, about 15,000 μg to about 25,000 μg, about 15,000 μg to about 20,000 μg, about 20,000 μg to about 100,000 μg, about 20,000 μg to Approximately 90,000 μg, approximately 20,000 μg to approximately 80,000 μg, approximately 20,000 μg to approximately 70,000 μg, approximately 20,000 μg to approximately 60,000 μg, approximately 20,000 μg to approximately 50,000 μg, approximately 20,000 μg to approximately 40,000 μg, approximately 20,000 μg to approximately 30,000 μg, approximately 20,000 μg to approximately 25,000 μg, approximately 25,000 μg to approximately 100,000 μg, approximately 25,000 μg to approximately 90,000 μg, approximately 25,000 μg to approximately 80,000 μg, approximately 25,000 μg to approximately 70,000 μg, approximately 25,000 μg to approximately 60,000 μg, approximately 25,000 μg to approximately 50,000 μg, approximately 2500 μg 0 μg to about 40,000 μg, about 25,000 μg to about 30,000 μg, about 30,000 μg to about 100,000 μg, about 30,000 μg to about 90,000 μg, about 30,000 μg to about 80,000 μg, about 30,000 μg to about 70,000 μg, about 30,000 μg to about 60,000 μg, about 30,000 μg to about 50,000 μg, about 30,000 μg to about 40,000 μg, about 40,000 μg to about 100,000 μg, about 40,000 μg to about 80,000 μg, about 40,000 μg to about 60,000 μg, about 40,000 μg to about 50,000 μg, about 50,000 μg to about 100,000 μg.Approximately 50,000 μg to approximately 70,000 μg, approximately 60,000 μg to approximately 100,000 μg, or approximately 70,000 μg to approximately 100,000 μg.

[0196] In some embodiments, the dose of the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) described herein is from about 1.0 μg to about 10,000 μg. In some embodiments, the dose is from about 1.0 μg to about 5,000 μg. In some embodiments, the dose is from about 1.0 μg to about 2,500 μg. In some embodiments, the dose is from about 1.0 μg to about 1,000 μg. In some embodiments, the dose is from about 1.0 μg to about 500 μg. In some embodiments, the dose is from about 1.0 μg to about 250 μg. In some embodiments, the dose is from about 1.0 μg to about 100 μg. In some embodiments, the dose is from about 1.0 μg to about 75 μg. In some embodiments, the dose is from about 1.0 μg to about 50 μg. In some embodiments, the dose is from about 1.0 μg to about 25 μg. In some embodiments, the dose is from about 1.0 μg to about 10 μg. In some embodiments, the dose is from about 10 μg to about 10,000 μg. In some embodiments, the dose is from about 10 μg to about 5,000 μg. In some embodiments, the dose is from about 10 μg to about 2,500 μg. In some embodiments, the dose is from about 10 μg to about 1,000 μg. In some embodiments, the dose is from about 10 μg to about 500 μg. In some embodiments, the dose is from about 10 μg to about 250 μg. In some embodiments, the dose is from about 10 μg to about 100 μg. In some embodiments, the dose is from about 10 μg to about 75 μg. In some embodiments, the dose is from about 10 μg to about 50 μg. In some embodiments, the dose is from about 50 μg to about 10,000 μg. In some embodiments, the dose is from about 50 μg to about 5,000 μg. In some embodiments, the dose is from about 50 μg to about 2,500 μg. In some embodiments, the dose is from about 50 μg to about 2,000 μg. In some embodiments, the dose is about 50 μg to about 1000 μg. In some embodiments, the dose is about 50 μg to about 750 μg. In some embodiments, the dose is about 50 μg to about 500 μg. In some embodiments, the dose is about 50 μg to about 250 μg. In some embodiments, the dose is about 50 μg to about 200 μg. In some embodiments, the dose is about 50 μg to about 100 μg. In some embodiments, the dose is about 100 μg to about 10000 μg. In some embodiments, the dose is about 100 μg to about 5000 μg. In some embodiments, the dose is about 100 μg to about 2500 μg. In some embodiments, the dose is about 100 μg to about 2000 μg. In some embodiments, the dose is about 100 μg to about 1500 μg. In some embodiments, the dose is about 100 μg to about 1250 μg. In some implementations, the dosage is from about 100 μg to about 1000 μg.In some embodiments, the dose is from about 100 μg to about 750 μg. In some embodiments, the dose is from about 100 μg to about 500 μg. In some embodiments, the dose is from about 100 μg to about 250 μg. In some embodiments, the dose is from about 100 μg to about 200 μg. In some embodiments, the dose is from about 250 μg to about 10000 μg. In some embodiments, the dose is from about 250 μg to about 5000 μg. In some embodiments, the dose is from about 250 μg to about 2500 μg. In some embodiments, the dose is from about 250 μg to about 2000 μg. In some embodiments, the dose is from about 250 μg to about 1500 μg. In some embodiments, the dose is from about 250 μg to about 1250 μg. In some embodiments, the dose is from about 250 μg to about 1000 μg. In some embodiments, the dose is from about 250 μg to about 750 μg. In some embodiments, the dose is from about 250 μg to about 500 μg. In some embodiments, the dose is from about 500 μg to about 10000 μg. In some embodiments, the dose is from about 500 μg to about 7000 μg. In some embodiments, the dose is from about 500 μg to about 5000 μg. In some embodiments, the dose is from about 500 μg to about 3000 μg. In some embodiments, the dose is from about 500 μg to about 2500 μg. In some embodiments, the dose is from about 500 μg to about 2000 μg. In some embodiments, the dose is from about 500 μg to about 1500 μg. In some embodiments, the dose is from about 500 μg to about 1250 μg. In some embodiments, the dose is from about 500 μg to about 1000 μg. In some embodiments, the dose is from about 500 μg to about 750 μg. In some embodiments, the dose is from about 750 μg to about 20000 μg. In some embodiments, the dose is about 750 μg to about 15000 μg. In some embodiments, the dose is about 750 μg to about 10000 μg. In some embodiments, the dose is about 750 μg to about 7000 μg. In some embodiments, the dose is about 750 μg to about 5000 μg. In some embodiments, the dose is about 750 μg to about 3000 μg. In some embodiments, the dose is about 750 μg to about 2500 μg. In some embodiments, the dose is about 750 μg to about 2000 μg. In some embodiments, the dose is about 750 μg to about 1500 μg. In some embodiments, the dose is about 750 μg to about 1250 μg. In some embodiments, the dose is about 750 μg to about 1000 μg. In some embodiments, the dose is about 1000 μg to about 25000 μg. In some implementations, the dosage is from about 1000 μg to about 20000 μg.In some embodiments, the dose is about 1000 μg to about 15000 μg. In some embodiments, the dose is about 1000 μg to about 10000 μg. In some embodiments, the dose is about 1000 μg to about 7000 μg. In some embodiments, the dose is about 1000 μg to about 5000 μg. In some embodiments, the dose is about 1000 μg to about 3000 μg. In some embodiments, the dose is about 1000 μg to about 2500 μg. In some embodiments, the dose is about 1000 μg to about 2000 μg. In some embodiments, the dose is about 1000 μg to about 1500 μg. In some embodiments, the dose is about 1500 μg to about 25000 μg. In some embodiments, the dose is about 1500 μg to about 20000 μg. In some embodiments, the dose is about 1500 μg to about 15000 μg. In some embodiments, the dose is from about 1500 μg to about 10000 μg. In some embodiments, the dose is from about 1500 μg to about 7000 μg. In some embodiments, the dose is from about 1500 μg to about 5000 μg. In some embodiments, the dose is from about 1500 μg to about 3000 μg. In some embodiments, the dose is from about 1500 μg to about 2500 μg. In some embodiments, the dose is from about 1500 μg to about 2000 μg. In some embodiments, the dose is from about 2000 μg to about 30000 μg. In some embodiments, the dose is from about 2000 μg to about 25000 μg. In some embodiments, the dose is from about 2000 μg to about 20000 μg. In some embodiments, the dose is from about 2000 μg to about 15000 μg. In some embodiments, the dose is from about 2000 μg to about 10000 μg. In some embodiments, the dose is from about 2000 μg to about 7000 μg. In some embodiments, the dose is from about 2000 μg to about 5000 μg. In some embodiments, the dose is from about 2000 μg to about 4000 μg. In some embodiments, the dose is from about 2000 μg to about 3000 μg. In some embodiments, the dose is from about 2000 μg to about 2500 μg. In some embodiments, the dose is from about 2500 μg to about 30000 μg. In some embodiments, the dose is from about 2500 μg to about 25000 μg. In some embodiments, the dose is from about 2500 μg to about 20000 μg. In some embodiments, the dose is from about 2500 μg to about 15000 μg. In some embodiments, the dose is from about 2500 μg to about 12000 μg. In some embodiments, the dose is from about 2500 μg to about 10000 μg. In some embodiments, the dosage is from about 2500 μg to about 7000 μg. In some embodiments, the dosage is from about 2500 μg to about 5000 μg.In some embodiments, the dose is about 2500 μg to about 4000 μg. In some embodiments, the dose is about 2500 μg to about 3000 μg. In some embodiments, the dose is about 3000 μg to about 50000 μg. In some embodiments, the dose is about 3000 μg to about 25000 μg. In some embodiments, the dose is about 3000 μg to about 20000 μg. In some embodiments, the dose is about 3000 μg to about 15000 μg. In some embodiments, the dose is about 3000 μg to about 12000 μg. In some embodiments, the dose is about 3000 μg to about 10000 μg. In some embodiments, the dose is about 3000 μg to about 7000 μg. In some embodiments, the dose is about 3000 μg to about 6000 μg. In some embodiments, the dose is about 3000 μg to about 5000 μg. In some embodiments, the dose is about 3000 μg to about 4000 μg. In some embodiments, the dose is about 4000 μg to about 50000 μg. In some embodiments, the dose is about 4000 μg to about 30000 μg. In some embodiments, the dose is about 4000 μg to about 25000 μg. In some embodiments, the dose is about 4000 μg to about 20000 μg. In some embodiments, the dose is about 4000 μg to about 15000 μg. In some embodiments, the dose is about 4000 μg to about 12000 μg. In some embodiments, the dose is about 4000 μg to about 10000 μg. In some embodiments, the dose is about 4000 μg to about 8000 μg. In some embodiments, the dose is about 4000 μg to about 6000 μg. In some embodiments, the dose is about 4000 μg to about 5000 μg. In some embodiments, the dose is about 5000 μg to about 50000 μg. In some embodiments, the dose is about 5000 μg to about 40000 μg. In some embodiments, the dose is about 5000 μg to about 30000 μg. In some embodiments, the dose is about 5000 μg to about 25000 μg. In some embodiments, the dose is about 5000 μg to about 20000 μg. In some embodiments, the dose is about 5000 μg to about 15000 μg. In some embodiments, the dose is about 5000 μg to about 12000 μg. In some embodiments, the dose is about 5000 μg to about 10000 μg. In some embodiments, the dose is about 5000 μg to about 8000 μg. In some embodiments, the dose is about 5000 μg to about 7000 μg. In some embodiments, the dose is about 5000 μg to about 6000 μg. In some implementations, the dosage is from about 6000 μg to about 50000 μg.In some embodiments, the dose is from about 6000 μg to about 30000 μg. In some embodiments, the dose is from about 6000 μg to about 25000 μg. In some embodiments, the dose is from about 6000 μg to about 20000 μg. In some embodiments, the dose is from about 6000 μg to about 17000 μg. In some embodiments, the dose is from about 6000 μg to about 15000 μg. In some embodiments, the dose is from about 6000 μg to about 12000 μg. In some embodiments, the dose is from about 6000 μg to about 10000 μg. In some embodiments, the dose is from about 6000 μg to about 9000 μg. In some embodiments, the dose is from about 6000 μg to about 8000 μg. In some embodiments, the dose is from about 6000 μg to about 7000 μg. In some embodiments, the dose is from about 7000 μg to about 70000 μg. In some embodiments, the dose is from about 7000 μg to about 50000 μg. In some embodiments, the dose is from about 7000 μg to about 40000 μg. In some embodiments, the dose is from about 7000 μg to about 30000 μg. In some embodiments, the dose is from about 7000 μg to about 25000 μg. In some embodiments, the dose is from about 7000 μg to about 20000 μg. In some embodiments, the dose is from about 7000 μg to about 17000 μg. In some embodiments, the dose is from about 7000 μg to about 15000 μg. In some embodiments, the dose is from about 7000 μg to about 12000 μg. In some embodiments, the dose is from about 7000 μg to about 10000 μg. In some embodiments, the dose is from about 7000 μg to about 8000 μg. In some embodiments, the dose is from about 8000 μg to about 80000 μg. In some embodiments, the dose is about 8000 μg to about 60000 μg. In some embodiments, the dose is about 8000 μg to about 50000 μg. In some embodiments, the dose is about 8000 μg to about 30000 μg. In some embodiments, the dose is about 8000 μg to about 25000 μg. In some embodiments, the dose is about 8000 μg to about 20000 μg. In some embodiments, the dose is about 8000 μg to about 17000 μg. In some embodiments, the dose is about 8000 μg to about 15000 μg. In some embodiments, the dose is about 8000 μg to about 12000 μg. In some embodiments, the dose is about 8000 μg to about 10000 μg. In some embodiments, the dose is about 10000 μg to about 80000 μg. In some implementations, the dosage is from about 10,000 μg to about 70,000 μg.In some embodiments, the dose is about 10,000 μg to about 60,000 μg. In some embodiments, the dose is about 10,000 μg to about 50,000 μg. In some embodiments, the dose is about 10,000 μg to about 40,000 μg. In some embodiments, the dose is about 10,000 μg to about 30,000 μg. In some embodiments, the dose is about 10,000 μg to about 25,000 μg. In some embodiments, the dose is about 10,000 μg to about 20,000 μg. In some embodiments, the dose is about 10,000 μg to about 18,000 μg. In some embodiments, the dose is about 10,000 μg to about 16,000 μg. In some embodiments, the dose is about 10,000 μg to about 15,000 μg. In some embodiments, the dose is about 10,000 μg to about 12,000 μg. In some embodiments, the dose is about 12,000 μg to about 100,000 μg. In some embodiments, the dose is about 12,000 μg to about 90,000 μg. In some embodiments, the dose is about 12,000 μg to about 80,000 μg. In some embodiments, the dose is about 12,000 μg to about 70,000 μg. In some embodiments, the dose is about 12,000 μg to about 60,000 μg. In some embodiments, the dose is about 12,000 μg to about 50,000 μg. In some embodiments, the dose is about 12,000 μg to about 40,000 μg. In some embodiments, the dose is about 12,000 μg to about 30,000 μg. In some embodiments, the dose is about 12,000 μg to about 25,000 μg. In some embodiments, the dose is about 12,000 μg to about 20,000 μg. In some embodiments, the dose is about 12,000 μg to about 18,000 μg. In some embodiments, the dose is about 12,000 μg to about 16,000 μg. In some embodiments, the dose is about 12,000 μg to about 15,000 μg. In some embodiments, the dose is about 12,000 μg to about 14,000 μg. In some embodiments, the dose is about 15,000 μg to about 100,000 μg. In some embodiments, the dose is about 15,000 μg to about 90,000 μg. In some embodiments, the dose is about 15,000 μg to about 80,000 μg. In some embodiments, the dose is about 15,000 μg to about 70,000 μg. In some embodiments, the dose is about 15,000 μg to about 60,000 μg. In some embodiments, the dose is about 15,000 μg to about 50,000 μg. In some embodiments, the dose is from about 15,000 μg to about 40,000 μg. In some embodiments, the dose is from about 15,000 μg to about 30,000 μg. In some embodiments, the dose is from about 15,000 μg to about 25,000 μg.In some embodiments, the dose is about 15,000 μg to about 20,000 μg. In some embodiments, the dose is about 20,000 μg to about 100,000 μg. In some embodiments, the dose is about 20,000 μg to about 90,000 μg. In some embodiments, the dose is about 20,000 μg to about 80,000 μg. In some embodiments, the dose is about 20,000 μg to about 70,000 μg. In some embodiments, the dose is about 20,000 μg to about 60,000 μg. In some embodiments, the dose is about 20,000 μg to about 50,000 μg. In some embodiments, the dose is about 20,000 μg to about 40,000 μg. In some embodiments, the dose is about 20,000 μg to about 30,000 μg. In some embodiments, the dose is about 20,000 μg to about 25,000 μg. In some embodiments, the dose is about 25,000 μg to about 100,000 μg. In some embodiments, the dose is about 25,000 μg to about 90,000 μg. In some embodiments, the dose is about 25,000 μg to about 80,000 μg. In some embodiments, the dose is about 25,000 μg to about 70,000 μg. In some embodiments, the dose is about 25,000 μg to about 60,000 μg. In some embodiments, the dose is about 25,000 μg to about 50,000 μg. In some embodiments, the dose is about 25,000 μg to about 40,000 μg. In some embodiments, the dose is about 25,000 μg to about 30,000 μg. In some embodiments, the dose is about 30,000 μg to about 100,000 μg. In some embodiments, the dose is about 30,000 μg to about 90,000 μg. In some embodiments, the dose is about 30,000 μg to about 80,000 μg. In some embodiments, the dose is about 30,000 μg to about 70,000 μg. In some embodiments, the dose is about 30,000 μg to about 60,000 μg. In some embodiments, the dose is about 30,000 μg to about 50,000 μg. In some embodiments, the dose is about 30,000 μg to about 40,000 μg. In some embodiments, the dose is about 40,000 μg to about 100,000 μg. In some embodiments, the dose is about 40,000 μg to about 80,000 μg. In some embodiments, the dose is about 40,000 μg to about 60,000 μg. In some embodiments, the dose is about 40,000 μg to about 50,000 μg. In some embodiments, the dose is about 50,000 μg to about 100,000 μg. In some embodiments, the dose is about 50,000 μg to about 70,000 μg. In some embodiments, the dose is about 60,000 μg to about 100,000 μg or about 70,000 μg to about 100,000 μg.

[0197] In some implementations, the dosage is about 5 μg, about 10 μg, about 25 μg, about 50 μg, about 75 μg, about 100 μg, about 125 μg, about 150 μg, about 175 μg, about 200 μg, about 225 μg, about 250 μg, about 275 μg, about 300 μg, about 325 μg, about 350 μg, about 375 μg, about 400 μg, about 425 μg, and about 450 μg. Approximately 475 μg, approximately 500 μg, approximately 550 μg, approximately 600 μg, approximately 650 μg, approximately 700 μg, approximately 750 μg, approximately 800 μg, approximately 850 μg, approximately 1000 μg, approximately 1250 μg, approximately 1500 μg, approximately 1750 μg, approximately 2000 μg, approximately 2500 μg, approximately 3000 μg, approximately 3500 μg, approximately 4000 μg, approximately 4500 μg, approximately 5 000μg, approximately 6000μg, approximately 7000μg, approximately 8000μg, approximately 9000μg, approximately 10000μg, approximately 11000μg, approximately 12000μg, approximately 13000μg, approximately 14000μg, approximately 15000μg, approximately 16000μg, approximately 17000μg, approximately 18000μg, approximately 19000μg, approximately 20000μg, approximately 21000μg Approximately 22,000 μg, approximately 23,000 μg, approximately 24,000 μg, approximately 25,000 μg, approximately 26,000 μg, approximately 27,000 μg, approximately 28,000 μg, approximately 29,000 μg, approximately 30,000 μg, approximately 40,000 μg, approximately 50,000 μg, approximately 60,000 μg, approximately 70,000 μg, approximately 80,000 μg, approximately 90,000 μg, or approximately 100,000 μg.

[0198] In some implementations, the dosage is approximately 100 μg, approximately 150 μg, approximately 300 μg, approximately 500 μg, approximately 750 μg, approximately 1000 μg, approximately 1500 μg, approximately 2500 μg, approximately 300 μg, approximately 3000 μg, approximately 3500 μg, approximately 4000 μg, approximately 4500 μg, approximately 5000 μg, approximately 6000 μg, approximately 7000 μg, approximately 8000 μg, approximately 9000 μg, approximately 10000 μg, approximately 11000 μg, approximately 12000 μg, approximately... 13000μg, approximately 14000μg, approximately 15000μg, approximately 16000μg, approximately 17000μg, approximately 18000μg, approximately 19000μg, approximately 20000μg, approximately 21000μg, approximately 22000μg, approximately 23000μg, approximately 24000μg, approximately 25000μg, approximately 26000μg, approximately 27000μg, approximately 28000μg, approximately 29000μg, approximately 30000μg, approximately 40000μg, or approximately 50000μg.

[0199] In some embodiments, the dosage is about 300 μg, about 300 μg, about 750 μg, about 1000 μg, about 1500 μg, about 2000 μg, about 2500 μg, about 3000 μg, about 4000 μg, or about 5000 μg. In some embodiments, the dosage is about 100 μg, about 150 μg, about 300 μg, about 750 μg, about 1000 μg, about 1500 μg, about 2000 μg, about 3000 μg, about 3500 μg, about 4000 μg, about 4500 μg, about 5000 μg, about 6000 μg, about 7000 μg, about 8000 μg, about 9000 μg, or about 1000 μg. 00 μg, approximately 11000 μg, approximately 12000 μg, approximately 13000 μg, approximately 14000 μg, approximately 15000 μg, approximately 16000 μg, approximately 17000 μg, approximately 18000 μg, approximately 19000 μg, approximately 20000 μg, approximately 21000 μg, approximately 22000 μg, approximately 23000 μg, approximately 24000 μg, or approximately 25000 μg.

[0200] In some implementations, the dosage is approximately 100 μg, approximately 150 μg, approximately 300 μg, approximately 750 μg, approximately 1000 μg, approximately 1500 μg, approximately 2000 μg, approximately 3000 μg, approximately 3500 μg, approximately 4000 μg, approximately 4500 μg, approximately 5000 μg, approximately 6000 μg, approximately 7000 μg, approximately 8000 μg, approximately 9000 μg, approximately 10000 μg, approximately 11000 μg, approximately 12000 μg, or approximately 13000 μg. Approximately 14,000 μg, approximately 15,000 μg, approximately 16,000 μg, approximately 17,000 μg, approximately 18,000 μg, approximately 19,000 μg, approximately 20,000 μg, approximately 21,000 μg, approximately 22,000 μg, approximately 23,000 μg, approximately 24,000 μg, approximately 25,000 μg, approximately 26,000 μg, approximately 27,000 μg, approximately 28,000 μg, approximately 29,000 μg, approximately 30,000 μg, approximately 40,000 μg, or approximately 50,000 μg.

[0201] In some implementations, the dosage is about 2000 μg, about 2500 μg, about 3000 μg, about 3500 μg, about 4000 μg, about 4500 μg, about 5000 μg, about 6000 μg, about 7000 μg, about 8000 μg, about 9000 μg, about 10000 μg, about 11000 μg, about 12000 μg, about 13000 μg, about 14000 μg, about 15000 μg, about 16000 μg, about 17000 μg, about 18000 μg, about 19000 μg, about 20000 μg, about 21000 μg, about 22000 μg, about 23000 μg, about 24000 μg, or about 25000 μg.

[0202] In some implementations, the dosage is about 2000 μg, about 2500 μg, about 3000 μg, about 3500 μg, about 4000 μg, about 4500 μg, about 5000 μg, about 6000 μg, about 7000 μg, about 8000 μg, about 9000 μg, or about 10000 μg.

[0203] In some implementations, the dosage is about 5000 μg, about 6000 μg, about 7000 μg, about 8000 μg, about 9000 μg, or about 10000 μg.

[0204] In some implementations, the dosage is about 7000 μg, about 8000 μg, about 9000 μg, or about 10000 μg, about 11000 μg, about 12000 μg, about 13000 μg, about 14000 μg, about 15000 μg, about 16000 μg, about 17000 μg, about 18000 μg, about 19000 μg, about 20000 μg, about 21000 μg, about 22000 μg, about 23000 μg, about 24000 μg, about 25000 μg, about 26000 μg, about 27000 μg, about 28000 μg, about 29000 μg, about 30000 μg, about 40000 μg, or about 50000 μg.

[0205] In some implementations, the dosage is about 100 μg, about 150 μg, about 300 μg, about 750 μg, about 1000 μg, about 1500 μg, about 2000 μg, about 25000 μg, about 3000 μg, about 4000 μg, or about 5000 μg.

[0206] In some embodiments, the dosage is about 10 μg, about 50 μg, about 100 μg, about 150 μg, about 300 μg, about 750 μg, or about 1000 μg. In some embodiments, the dosage is about 50 μg, about 100 μg, about 150 μg, about 300 μg, about 750 μg, or about 1000 μg. In some embodiments, the dosage is about 100 μg, about 150 μg, about 300 μg, about 750 μg, about 1000 μg, about 1500 μg, about 2000 μg, about 3000 μg, about 4000 μg, or about 5000 μg. In some embodiments, the dosage is about 100 μg, about 150 μg, about 300 μg, about 750 μg, about 1000 μg, about 1500 μg, about 2000 μg, or about 3000 μg. In some embodiments, the dosage is about 300 μg, about 750 μg, about 1000 μg, about 1500 μg, or about 2000 μg. In some embodiments, the dosage is about 150 μg, about 300 μg, about 750 μg, or about 1000 μg. In some embodiments, the dosage is about 300 μg, about 750 μg, or about 1000 μg. In some embodiments, the dosage is about 150 μg, about 300 μg, or about 750 μg. In some embodiments, the dosage is about 300 μg, about 750 μg, or about 1000 μg. In some embodiments, the dose is about 150 μg or about 300 μg. In some embodiments, the dose is about 300 μg or about 750 μg. In some embodiments, the dose is about 750 μg or about 1000 μg.

[0207] In some embodiments, the dose is about 5 μg. In some embodiments, the dose is about 10 μg. In some embodiments, the dose is about 25 μg. In some embodiments, the dose is about 50 μg. In some embodiments, the dose is about 75 μg. In some embodiments, the dose is about 100 μg. In some embodiments, the dose is about 125 μg. In some embodiments, the dose is about 150 μg. In some embodiments, the dose is about 175 μg. In some embodiments, the dose is about 200 μg. In some embodiments, the dose is about 225 μg. In some embodiments, the dose is about 250 μg. In some embodiments, the dose is about 275 μg. In some embodiments, the dose is about 300 μg. In some embodiments, the dose is about 325 μg. In some embodiments, the dose is about 350 μg. In some embodiments, the dose is about 375 μg. In some embodiments, the dose is about 400 μg. In some embodiments, the dose is about 425 μg. In some embodiments, the dose is about 450 μg. In some embodiments, the dose is about 475 μg. In some embodiments, the dose is about 500 μg. In some embodiments, the dose is about 550 μg. In some embodiments, the dose is about 600 μg. In some embodiments, the dose is about 650 μg. In some embodiments, the dose is about 700 μg. In some embodiments, the dose is about 750 μg. In some embodiments, the dose is about 800 μg. In some embodiments, the dose is about 850 μg. In some embodiments, the dose is about 1000 μg. In some embodiments, the dose is about 1250 μg. In some embodiments, the dose is about 1500 μg. In some embodiments, the dose is about 1750 μg. In some embodiments, the dose is about 2000 μg. In some embodiments, the dose is about 2500 μg. In some embodiments, the dose is about 3000 μg. In some embodiments, the dose is about 3500 μg. In some embodiments, the dose is about 4000 μg. In some embodiments, the dose is about 4500 μg. In some embodiments, the dose is about 5000 μg. In some embodiments, the dose is about 6000 μg. In some embodiments, the dose is about 7000 μg. In some embodiments, the dose is about 8000 μg. In some embodiments, the dose is about 9000 μg. In some embodiments, the dose is about 10000 μg. In some embodiments, the dose is about 11000 μg. In some embodiments, the dose is about 12000 μg. In some embodiments, the dose is about 13000 μg. In some embodiments, the dose is about 14000 μg. In some embodiments, the dose is about 15000 μg. In some embodiments, the dose is about 16000 μg.In some embodiments, the dose is about 17,000 μg. In some embodiments, the dose is about 18,000 μg. In some embodiments, the dose is about 19,000 μg. In some embodiments, the dose is about 20,000 μg. In some embodiments, the dose is about 21,000 μg. In some embodiments, the dose is about 22,000 μg. In some embodiments, the dose is about 23,000 μg. In some embodiments, the dose is about 24,000 μg. In some embodiments, the dose is about 25,000 μg. In some embodiments, the dose is about 26,000 μg. In some embodiments, the dose is about 27,000 μg. In some embodiments, the dose is about 28,000 μg. In some embodiments, the dose is about 29,000 μg. In some embodiments, the dose is about 30,000 μg. In some embodiments, the dose is about 40,000 μg. In some embodiments, the dose is about 50,000 μg. In some embodiments, the dose is about 60,000 μg. In some embodiments, the dose is about 70,000 μg. In some embodiments, the dose is about 80,000 μg. In some embodiments, the dose is about 90,000 μg. In some embodiments, the dose is about 100,000 μg.

[0208] In some embodiments, the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) is administered daily. In some embodiments, the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) is administered every other day. In some embodiments, the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) is administered every four days. In some embodiments, the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) is administered every three days. In some embodiments, the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) is administered every five days. In some embodiments, the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) is administered every six days. In some embodiments, the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) is administered weekly. In some embodiments, the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) is administered every ten days. In some embodiments, the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) is administered every two weeks. In some embodiments, the ETBR antagonist (e.g., a specific deuterated ETBR antagonist) is administered every three weeks.

[0209] Application route

[0210] This document discloses various routes of administration for the pharmaceutical compositions disclosed herein. According to this disclosure, compounds as described herein can be administered via oral, parenteral, or surface routes in a single dose or multiple doses. The range of administration for the active compound can range from continuous (intravenous infusion) to several oral administrations per day (e.g., QOD or QID), and may include oral, surface, parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include a penetration enhancer), sublingual, sublingual, and suppository administration, as well as other routes of administration. Enteric-coated oral tablets can also be used to enhance the bioavailability of the compound from the oral administration route. The most effective dosage form will depend on the pharmacokinetics of one or more specific agents selected and the severity of the patient's disease. The compounds according to this disclosure can also be used as sprays, mists, or aerosols for intranasal, intratracheal, or pulmonary administration. Therefore, this disclosure also relates to pharmaceutical compositions comprising an effective amount of the compound as described herein, optionally combined with a pharmaceutically acceptable carrier, additive, or excipient. The compounds according to this disclosure can be administered in an immediate-release, intermediate-release, sustained-release, or controlled-release form. In some embodiments, sustained-release or controlled-release forms are administered orally, but are also administered as suppositories and transdermal or other surface forms. Intramuscular injection in liposome form can also be used to control or sustain the release of the compound at the injection site.

[0211] In some embodiments, the pharmaceutical compositions described herein are administered orally, parenterally, by inhalation spray, surface, rectum, nose, sublingually, vaginally, or via an implanted reservoir. As used herein, the term "parentereal" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. In some embodiments, the compositions are administered orally, intraperitoneally, or intravenously.

[0212] In some embodiments, the sterile injectable form of the compositions described herein is an aqueous or oily suspension. These suspensions can be formulated using suitable dispersants or wetting agents and suspending agents. Sterile injectable articles can also be sterile injectable solutions or suspensions in non-toxic, parenteral-acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable media and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. Additionally, sterile non-volatile oils are conventionally used as solvents or suspension media. For this purpose, any mild non-volatile oil can be used, including synthetic monoglycerides or diglycerides. Fatty acids such as oleic acid and its glycerol derivatives that can be used to prepare injectables are pharmaceutically acceptable natural oils such as olive oil, castor oil, or soybean oil, especially their polyoxyethylated forms. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as Ph. Helv or similar alcohols.

[0213] In some embodiments, the pharmaceutical compositions described herein are administered orally in any orally acceptable dosage form, including but not limited to capsules, tablets, aqueous suspensions, or solutions. In the case of tablets for oral use, commonly used carriers include lactose and corn starch. Lubricants, such as magnesium stearate, are also typically added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. When used orally in aqueous suspensions, the active ingredient is combined with an emulsifier and a suspending agent. If desired, certain sweeteners, flavoring agents, or coloring agents may also be added.

[0214] In some embodiments, the pharmaceutical compositions described herein are administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and thus melts in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol.

[0215] In some embodiments, the pharmaceutical compositions described herein are applied topically. Suitable surface formulations are readily prepared for each of these areas or organs. Surface application for the lower intestine can be made in rectal suppository formulations (see above) or suitable enema formulations. Surface-acceptable transdermal patches may also be used.

[0216] In some embodiments, for surface application, the pharmaceutical composition is formulated in a suitable ointment containing an active ingredient suspended or dissolved in one or more carriers. Carriers for surface application of the compounds of this disclosure include, but are not limited to, mineral oil, liquid petrolatum, DMSO, white petrolatum, propylene glycol, polyethylene oxide, polyoxypropylene compounds, emulsified waxes, and water. In some embodiments, the compound may be coated onto a stent to be surgically implanted into a patient to inhibit or reduce the likelihood of stent occlusion within the patient.

[0217] In some embodiments, the pharmaceutical composition is formulated in a suitable lotion or cream containing an active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecyl alcohol, benzyl alcohol, and water.

[0218] In some embodiments, for ophthalmic use, the pharmaceutical composition is formulated as a micronized suspension in isotonic, pH-adjusted sterile saline, or as a solution in isotonic, pH-adjusted sterile saline, with or without a preservative such as benzalkonium chloride. Optionally, for ophthalmic use, the pharmaceutical composition may be formulated in an ointment such as petrolatum.

[0219] In some embodiments, the pharmaceutical compositions described herein are administered via nasal aerosol or inhalation. Such compositions are prepared according to the techniques described herein relating to pharmaceutical compositions and can be prepared as solutions in saline using benzyl alcohol or other suitable preservatives, bioavailability-enhancing absorption promoters, fluorocarbons, and / or other conventional solubilizers or dispersants. In some embodiments, this specification provides formulations comprising liposomes containing an effective amount (e.g., a synergistically effective amount) of an ETBR antagonist.

[0220] In some embodiments, the composition is formulated to contain an active ingredient, alone or in combination with another compound according to this disclosure, in amounts of about 0.05 mg to about 750 mg or more, such as about 1 mg to about 600 mg, or about 10 mg to about 500 mg. It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the particular compound used, age, weight, general health, sex, diet, time of administration, excretion rate, drug combination, and the judgment of the treating physician and the severity of the particular disease or condition being treated.

[0221] In some implementations, a patient or subject requiring therapy with a compound according to the methods described herein is treated by administering to the patient (subject) an effective amount of a compound, alone or in combination with other known erythropoiesis stimulants as otherwise identified herein, including optionally a pharmaceutically acceptable salt, solvate, or polymorph thereof in a pharmaceutically acceptable carrier or diluent.

[0222] In some embodiments, the compounds or compositions herein are administered orally, parenterally, intradermally, by injection (intravenous, subcutaneous, or intramuscular), or topically (including transdermal) in liquid, cream, gel, or solid form or by aerosol.

[0223] In some embodiments, the active ingredient is contained in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver a therapeutically effective amount to a patient for the desired indication without causing serious toxicity in the treated patient. Exemplary doses of the active compound for all conditions mentioned herein are in the range of about 10 ng / kg to 300 ng / kg, about 10 ng / kg to 1 μg / kg, about 1 μg / kg to 10 μg / kg, about 10 μg / kg to 100 μg / kg, about 100 μg / kg to 1000 μg / kg, about 1 mg / kg to 30 mg / kg, about 1 mg / kg to 300 mg / kg, or 0.1 mg / kg to 100 mg / kg daily, and more typically 0.5 mg / kg to about 25 mg / kg of recipient / patient body weight daily. Typical surface doses will be in the range of 0.01% to 5% wt / wt in a suitable carrier.

[0224] In some embodiments, the active ingredient herein is conveniently administered in any suitable unit dosage form, including but not limited to unit dosage forms containing less than 1 μg, or 1 μg to 5000 μg, for example, 50 μg to 3000 μg of active ingredient per unit dosage form.

[0225] In some embodiments, the active ingredient is administered to achieve peak plasma concentrations of the active compound at approximately 0.00001 mM to 30 mM, for example, approximately 0.1 μM to 30 μM. This can be achieved, for example, by intravenous injection of a solution or formulation of the active ingredient (optionally in a saline or aqueous medium) or by bolus administration of the active ingredient. Oral administration is also suitable for producing effective plasma concentrations of the active agent.

[0226] Immune cells expressing chimeric antigen receptors

[0227] The combination therapies and methods described herein typically involve immune cells expressing chimeric antigen receptors (CARs). A chimeric antigen receptor (CAR) is a synthetic receptor that mimics immune receptor activation to direct immune cell activity against a target of interest. A CAR essentially consists of: (a) an extracellular domain that binds to the target of interest (e.g., a single-chain variable fragment (scFv)), (b) a transmembrane domain, and (c) an intracellular domain that activates immune signaling (e.g., a CD3z domain). CARs can be expressed in a variety of different types of immune cells that can be used in the methods described herein. In some embodiments, CARs are expressed in T cells (e.g., CD4 T cells, CD8 T cells, T...). C 1 cell, T C 2 cells, γδT cells, T cells H 1 cell, T H 2 cells, T H 3. T H 17 cells, T FH Cells, T reg CAR is expressed in T cells (including memory T cells, NK T cells, and NK cells) or NK cells. In some embodiments, CAR is expressed in T cells (“CAR-T cells”). In some embodiments, CAR is expressed in NK cells (“CAR-NK cells”).

[0228] In some embodiments, the CAR used in the methods described herein can specifically bind to tumor-associated antigens. "Tumor-associated antigens" are proteins produced by tumor cells that trigger an immune response. Tumor-associated antigens include, for example, human epidermal growth factor receptor 2 (HER2). In some embodiments, the tumor-associated antigen is human epidermal growth factor receptor 2 (HER2).

[0229] In some implementations, the dose of CAR-expressing immune cells is 10. 4 One to 10 9 Cells / kg body weight, 10 5 One to 10 9 Cells / kg body weight, 10 6 One to 10 9 Cells / kg body weight, 10 7 One to 10 9 Cells / kg body weight, 10 8 One to 10 9 Cells / kg body weight, 10 4 One to 10 8 Cells / kg body weight, 10 5 One to 10 8 Cells / kg body weight, 10 6 One to 10 8 Cells / kg body weight, 10 7 One to 10 8 Cells / kg body weight, 10 4 One to 10 7 Cells / kg body weight, 10 5 One to 10 7 Cells / kg body weight, 10 6 One to 10 7 Cells / kg body weight, 10 4 One to 10 6 Cells / kg body weight, 10 5 One to 10 6 Cells / kg body weight or 10 4 One to 10 5 Cells per kg of body weight.

[0230] In some embodiments, CAR-expressing immune cells are administered in a single infusion. In some embodiments, CAR-expressing immune cells are administered in multiple infusions (e.g., 2-20 infusions). In some embodiments, CAR-expressing immune cells are administered in two infusions. In some embodiments, CAR-expressing immune cells are administered in three infusions. In some embodiments, CAR-expressing immune cells are administered in four infusions. In some embodiments, CAR-expressing immune cells are administered in five infusions. In some embodiments, CAR-expressing immune cells are administered in six infusions. In some embodiments, CAR-expressing immune cells are administered in seven infusions. In some embodiments, CAR-expressing immune cells are administered in eight infusions. In some embodiments, CAR-expressing immune cells are administered in nine infusions. In some embodiments, CAR-expressing immune cells are administered in ten infusions. In some embodiments, CAR-expressing immune cells are administered in eleven infusions. In some embodiments, CAR-expressing immune cells are administered in twelve infusions. In some embodiments, CAR-expressing immune cells are administered in thirteen infusions. In some embodiments, CAR-expressing immune cells are administered in fourteen infusions. In some embodiments, CAR-expressing immune cells are administered in sixteen infusions. In some embodiments, CAR-expressing immune cells are administered in seventeen infusions. In some embodiments, CAR-expressing immune cells are administered in eighteen infusions. In some embodiments, CAR-expressing immune cells are administered in nineteen infusions. In some embodiments, CAR-expressing immune cells are administered in twenty infusions.

[0231] In some embodiments, CAR-expressing immune cells are administered at weekly intervals. In some embodiments, CAR-expressing immune cells are administered at two-week intervals. In some embodiments, CAR-expressing immune cells are administered at three-week intervals. In some embodiments, CAR-expressing immune cells are administered at four-week intervals. In some embodiments, CAR-expressing immune cells are administered at five-week intervals. In some embodiments, CAR-expressing immune cells are administered at six-week intervals. In some embodiments, CAR-expressing immune cells are administered at seven-week intervals. In some embodiments, CAR-expressing immune cells are administered at eight-week intervals. In some embodiments, CAR-expressing immune cells are administered at nine-week intervals. In some embodiments, CAR-expressing immune cells are administered at ten-week intervals. In some embodiments, CAR-expressing immune cells are administered at eleven-week intervals. In some embodiments, CAR-expressing immune cells are administered at twelve-week intervals.

[0232] In some embodiments, CAR-expressing immune cells are administered at monthly intervals. In some embodiments, CAR-expressing immune cells are administered at two-month intervals. In some embodiments, CAR-expressing immune cells are administered at three-month intervals. In some embodiments, CAR-expressing immune cells are administered at four-month intervals. In some embodiments, CAR-expressing immune cells are administered at five-month intervals. In some embodiments, CAR-expressing immune cells are administered at six-month intervals. In some embodiments, CAR-expressing immune cells are administered at seven-month intervals. In some embodiments, CAR-expressing immune cells are administered at eight-month intervals. In some embodiments, CAR-expressing immune cells are administered at nine-month intervals. In some embodiments, CAR-expressing immune cells are administered at ten-month intervals. In some embodiments, CAR-expressing immune cells are administered at eleven-month intervals. In some embodiments, CAR-expressing immune cells are administered at twelve-month intervals.

[0233] Immune checkpoint inhibitors

[0234] In various embodiments, the methods disclosed herein include administering a combination therapy comprising an immune checkpoint inhibitor (ICPI) to a subject. In some embodiments, the ICPI is an anti-PD1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-LAG-3 antibody, or a CISH inhibitor. In some embodiments, the ICPI is an anti-PD1 antibody. In some embodiments, the anti-PD1 antibody is nivolumab, pembrolizumab, pidilizumab, or any combination thereof. In some embodiments, the anti-PD1 antibody is nivolumab. In some embodiments, the anti-PD1 antibody is pembrolizumab. In some embodiments, the anti-PD1 antibody is pidilizumab. In some embodiments, the anti-PD1 antibody is an anti-PD-L1 antibody. In some embodiments, the anti-PD-L1 antibody is atezolizumab, MDX-1105, avelumab, durvalumab, or any combination thereof. In some embodiments, the anti-PD-L1 antibody is atezolizumab. In some embodiments, the anti-PD-L1 antibody is MDX-1105. In some embodiments, the anti-PD-L1 antibody is avelumab. In some embodiments, the anti-PD-L1 antibody is durvalumab.

[0235] In some implementations, the dosage of the ICPI is from about 0.1 mg / kg to about 9.0 mg / kg. For example, the dosage of the anti-PD1 antibody is from about 0.1 mg / kg to about 9.0 mg / kg, from about 0.1 mg / kg to about 8.0 mg / kg, from about 0.1 mg / kg to about 7.0 mg / kg, from about 0.1 mg / kg to about 6.0 mg / kg, from about 0.1 mg / kg to about 5.0 mg / kg, from about 0.1 mg / kg to about 4.0 mg / kg, from about 0.1 mg / kg to about 3.0 mg / kg, from about 0.1 mg / kg to about 2.0 mg / kg, from about 0.1 mg / kg to about 1.0 mg / kg, from about 1.0 mg / kg to about 9.0 mg / kg, and from about 1.0 mg / kg to about 8.0 mg / kg. g / kg, about 1.0 mg / kg to about 7.0 mg / kg, about 1.0 mg / kg to about 6.0 mg / kg, about 1.0 mg / kg to about 5.0 mg / kg, about 1.0 mg / kg to about 4.0 mg / kg, about 1.0 mg / kg to about 3.0 mg / kg, about 1.0 mg / kg to about 2.0 mg / kg, about 2.0 mg / kg to about 9.0 mg / kg, about 2.0 mg / kg to about 8.0 mg / kg, about 2.0 mg / kg to about 7.0 mg / kg, about 2.0 mg / kg to about 6.0 mg / kg, about 2.0 mg / kg to about 5.0 mg / kg, about 2. 0 mg / kg to about 4.0 mg / kg, about 2.0 mg / kg to about 3.0 mg / kg, about 3.0 mg / kg to about 9.0 mg / kg, about 3.0 mg / kg to about 8.0 mg / kg, about 3.0 mg / kg to about 7.0 mg / kg, about 3.0 mg / kg to about 6.0 mg / kg, about 3.0 mg / kg to about 5.0 mg / kg, about 3.0 mg / kg to about 4.0 mg / kg, about 4.0 mg / kg to about 9.0 mg / kg, about 4.0 mg / kg to about 8.0 mg / kg, about 4.0 mg / kg to about 7.0 mg / kg, about 4.0 mg / kg to about 6.0 mg / kg, about 4.0 mg / kg to about 5.0 mg / kg, about 5.0 mg / kg to about 9.0 mg / kg, about 5.0 mg / kg to about 8.0 mg / kg, about 5.0 mg / kg to about 7.0 mg / kg, about 5.0 mg / kg to about 6.0 mg / kg, about 6.0 mg / kg to about 9.0 mg / kg, about 6.0 mg / kg to about 8.0 mg / kg, about 6.0 mg / kg to about 7.0 mg / kg, about 7.0 mg / kg to about 9.0 mg / kg, about 7.0 mg / kg to about 8.0 mg / kg, or about 8.0 mg / kg to about 9.0 mg / kg.

[0236] In some embodiments, the dosage of the ICPI is from about 10 mg to about 5000 mg. In some embodiments, the dosage of the ICPI is from about 10 mg to about 900 mg. For example, the dosage of the anti-PD1 antibody is from about 10 mg to about 900 mg, from about 10 mg to about 800 mg, from about 10 mg to about 700 mg, from about 10 mg to about 600 mg, from about 10 mg to about 500 mg, from about 10 mg to about 400 mg, from about 10 mg to about 300 mg, from about 10 mg to about 200 mg, from about 10 mg to about 100 mg, from about 100 mg to about 900 mg, from about 100 mg to about 800 mg. About 100mg to about 700mg, about 100mg to about 600mg, about 100mg to about 500mg, about 100mg to about 400mg, about 100mg to about 300mg, about 100mg to about 200mg, about 200mg to about 900mg, about 200mg to about 800mg, about 200mg to about 700mg, about 200mg to about 600mg, about 200mg to about 500mg, about 200mg g to about 400mg, about 200mg to about 300mg, about 300mg to about 900mg, about 300mg to about 800mg, about 300mg to about 700mg, about 300mg to about 600mg, about 300mg to about 500mg, about 300mg to about 400mg, about 400mg to about 900mg, about 400mg to about 800mg, about 400mg to about 700mg, about 400mg to about 6 00mg, about 400mg to about 500mg, about 500mg to about 900mg, about 500mg to about 800mg, about 500mg to about 700mg, about 500mg to about 600mg, about 600mg to about 900mg, about 600mg to about 800mg, about 600mg to about 700mg, about 700mg to about 900mg, about 700mg to about 800mg or about 800mg to about 900mg.

[0237] In some implementations, the dosage of ICPI is approximately 10 mg, approximately 20 mg, approximately 30 mg, approximately 40 mg, approximately 50 mg, approximately 70 mg, approximately 75 mg, approximately 80 mg, approximately 90 mg, approximately 100 mg, approximately 150 mg, approximately 200 mg, approximately 250 mg, approximately 300 mg, approximately 350 mg, approximately 400 mg, approximately 450 mg, approximately 500 mg, approximately 550 mg, approximately 600 mg, approximately 650 mg, approximately 700 mg, approximately 750 mg. mg, approximately 800mg, approximately 850mg, approximately 900mg, approximately 950mg, approximately 1000mg, approximately 1100mg, approximately 1200mg, approximately 1300mg, approximately 1400mg, approximately 1500mg, approximately 1600mg, approximately 1700mg, approximately 1800mg, approximately 1900mg, approximately 2000mg, approximately 2500mg, approximately 3000mg, approximately 3500mg, approximately 4000mg, approximately 4500mg, or approximately 5000mg. In some implementations, the dosage of ICPI is about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 70 mg, about 75 mg, about 80 mg, about 90 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg.

[0238] In some embodiments, the ICPI dose is about 10 mg. In some embodiments, the ICPI dose is about 20 mg. In some embodiments, the ICPI dose is about 30 mg. In some embodiments, the ICPI dose is about 40 mg. In some embodiments, the ICPI dose is about 50 mg. In some embodiments, the ICPI dose is about 70 mg. In some embodiments, the ICPI dose is about 75 mg. In some embodiments, the ICPI dose is about 80 mg. In some embodiments, the ICPI dose is about 90 mg. In some embodiments, the ICPI dose is about 100 mg. In some embodiments, the ICPI dose is about 150 mg. In some embodiments, the ICPI dose is about 200 mg. In some embodiments, the ICPI dose is about 250 mg. In some embodiments, the ICPI dose is about 300 mg. In some embodiments, the ICPI dose is about 350 mg. In some embodiments, the ICPI dose is about 400 mg. In some embodiments, the ICPI dose is about 450 mg. In some embodiments, the ICPI dose is about 500 mg. In some embodiments, the ICPI dose is about 550 mg. In some embodiments, the ICPI dose is about 600 mg. In some embodiments, the ICPI dose is about 650 mg. In some embodiments, the ICPI dose is about 700 mg. In some embodiments, the ICPI dose is about 750 mg. In some embodiments, the ICPI dose is about 800 mg. In some embodiments, the ICPI dose is about 850 mg. In some embodiments, the ICPI dose is about 900 mg. In some embodiments, the ICPI dose is about 950 mg. In some embodiments, the ICPI dose is about 1000 mg. In some embodiments, the ICPI dose is about 1100 mg. In some embodiments, the ICPI dose is about 1200 mg. In some embodiments, the ICPI dose is about 1300 mg. In some embodiments, the ICPI dose is about 1400 mg. In some embodiments, the ICPI dose is about 1500 mg. In some embodiments, the ICPI dose is about 1600 mg. In some embodiments, the ICPI dose is about 1700 mg. In some embodiments, the ICPI dose is about 1800 mg. In some embodiments, the ICPI dose is about 1900 mg. In some embodiments, the ICPI dose is about 2000 mg. In some embodiments, the ICPI dose is about 2500 mg. In some embodiments, the ICPI dose is about 3000 mg. In some embodiments, the ICPI dose is about 3500 mg.In some embodiments, the ICPI dose is approximately 4000 mg. In some embodiments, the ICPI dose is approximately 4500 mg. In some embodiments, the ICPI dose is approximately 5000 mg.

[0239] In some embodiments, ICPI is administered daily. In some embodiments, ICPI is administered every other day. In some embodiments, ICPI is administered every three days. In some embodiments, ICPI is administered every four days. In some embodiments, ICPI is administered every five days. In some embodiments, ICPI is administered every six days. In some embodiments, ICPI is administered every seven days. In some embodiments, ICPI is administered every eight days. In some embodiments, ICPI is administered every nine days. In some embodiments, ICPI is administered every ten days. In some embodiments, ICPI is administered every eleven days. In some embodiments, ICPI is administered every twelve days. In some embodiments, ICPI is administered every thirteen days. In some embodiments, ICPI is administered every fourteen days. In some embodiments, ICPI is administered every fifteen days. In some embodiments, ICPI is administered every sixteen days. In some embodiments, ICPI is administered every seventeen days. In some embodiments, ICPI is administered every eighteen days. In some embodiments, ICPI is administered every nineteen days. In some embodiments, ICPI is administered every twenty days. In some implementations, ICPI is administered every 21 days. In some implementations, ICPI is administered weekly. In some implementations, ICPI is administered every two weeks. In some implementations, ICPI is administered every three weeks. In some implementations, ICPI is administered every four weeks. In some implementations, ICPI is administered every five weeks. In some implementations, ICPI is administered every six weeks. In some implementations, ICPI is administered every seven weeks. In some implementations, ICPI is administered every eight weeks. In some implementations, ICPI is administered every nine weeks. In some implementations, ICPI is administered every ten weeks. In some implementations, ICPI is administered every eleven weeks. In some implementations, ICPI is administered every twelve weeks.

[0240] Reagent test kit

[0241] This document discloses kits or pharmaceutical compositions for treating solid tumor cancers in subjects (e.g., human subjects), said kits or pharmaceutical compositions comprising an effective amount of an ETBR antagonist for use in combination therapy with CAR-expressing immune cells and a pharmaceutically acceptable carrier. In some embodiments, the kits or pharmaceutical compositions also comprise an ICPI (e.g., an anti-PD1 antibody). In some embodiments, the ETBR antagonist is a specific deuterated ETBR antagonist as described herein, such as deuterated BQ-788. In some embodiments, the ETBR antagonist (e.g., deuterated BQ-788) and CAR-expressing immune cells are disposed in a single container. In some embodiments, the ETBR antagonist (e.g., deuterated BQ-788) is disposed in a first container and the CAR-expressing immune cells are disposed in a second container, wherein the ETBR antagonist and the CAR-expressing immune cells are administered approximately simultaneously.

[0242] In some embodiments, this specification provides a kit for treating solid tumor cancer in human subjects, the kit comprising a CAR-expressing immune cells, a synergistically effective amount of an ETBR antagonist (e.g., a specific deuterated ETBR antagonist as described herein), and a pharmaceutically acceptable carrier or excipient. In some embodiments, the kit or pharmaceutical composition further comprises an ICPI (e.g., an anti-PD1 antibody). In some embodiments, the ETBR antagonist is deuterated BQ-788. In some embodiments, the ETBR antagonist is BQ-788-B (ENB-003). In some embodiments, the ICPI is an anti-PD1 antibody or an anti-PD-L1 antibody.

[0243] Treatment

[0244] This document discloses methods for treating or improving cancer in subjects or patients (e.g., animals such as humans), methods comprising administering an effective amount (e.g., a therapeutically effective amount or a synergistically effective amount) of a combination therapy as described herein to a subject with an appropriate need, wherein the combination therapy is effective in treating or improving the subject's disease or disorder or its symptoms. In some embodiments, the combination therapy comprises CAR-expressing immune cells and an ETBR antagonist (e.g., a deuterated ETBR antagonist). In some embodiments, the combination therapy comprises CAR-expressing immune cells, an ICPI, and an ETBR antagonist (e.g., a deuterated ETBR antagonist). In some implementations, the ETBR antagonist (e.g., a deuterated ETBR antagonist) is administered 2, 3, 4, or 5 times more frequently than the CAR-expressing immune cells or ICPI. For example, the ETBR antagonist is administered 3 times over a 1-3 week period (e.g., over a period of about 2-3 weeks or about 21 days), while the CAR-expressing immune cells are administered 1 or 2 times over a 1-3 week period (e.g., over a period of about 2-3 weeks or about 21 days).

[0245] In some embodiments, the disease or disorder is cancer expressing the target. In some embodiments, the cancer is refractory or insensitive to immunotherapy (e.g., with checkpoint inhibitors and / or with CAR-expressing immunotherapy, such as CAR-T therapy) or both. In some embodiments, the cancer is newly diagnosed. In some embodiments, the cancer is relapsed or refractory. In some embodiments, the cancer is relapsed. In some embodiments, the cancer is refractory.

[0246] In some implementations, the cancer is ETBR-associated cancer. In some implementations, the cancer is HER2-expressing cancer. In some implementations, the cancer is HER2+ cancer. In some implementations, the cancer is low-HER2 cancer. In some implementations, the cancer is a hematologic malignancy. In some implementations, the cancer is a solid tumor. In some implementations, the cancer is selected from malignant melanoma, malignant squamous cell carcinoma, glioblastoma, pancreatic cancer, colon cancer, breast cancer, ovarian cancer, prostate cancer, bladder cancer, and kidney cancer. In some implementations, the cancer is selected from breast cancer, stomach cancer, esophageal cancer, ovarian cancer, endometrial cancer, bladder cancer, lung cancer, cervical cancer, colon cancer, and head and neck cancer. In some implementations, the cancer is breast cancer. In some implementations, the cancer is stomach cancer. In some implementations, the cancer is esophageal cancer. In some implementations, the cancer is ovarian cancer. In some implementations, the cancer is endometrial cancer. In some implementations, the cancer is bladder cancer. In some implementations, the cancer is lung cancer. In some implementations, the cancer is cervical cancer. In some implementations, the cancer is colon cancer. In some implementations, the cancer is head and neck cancer.

[0247] In some implementations, the cancer is HER2+ breast cancer, HER2+ stomach cancer, HER2+ esophageal cancer, HER2+ ovarian cancer, HER2+ endometrial cancer, HER2+ bladder cancer, HER2+ lung cancer, HER2+ cervical cancer, HER2+ colon cancer, or HER2+ head and neck cancer. In some implementations, the cancer is HER2+ breast cancer. In some implementations, the cancer is HER2+ stomach cancer. In some implementations, the cancer is HER2+ esophageal cancer. In some implementations, the cancer is HER2+ ovarian cancer. In some implementations, the cancer is HER2+ endometrial cancer. In some implementations, the cancer is HER2+ bladder cancer. In some implementations, the cancer is HER2+ lung cancer. In some implementations, the cancer is HER2+ cervical cancer. In some implementations, the cancer is HER2+ colon cancer. In some implementations, the cancer is HER2+ head and neck cancer.

[0248] In some embodiments, the cancer is low-HER2 breast cancer, low-HER2 gastric cancer, low-HER2 esophageal cancer, low-HER2 ovarian cancer, low-HER2 endometrial cancer, low-HER2 bladder cancer, low-HER2 lung cancer, low-HER2 cervical cancer, low-HER2 colon cancer, or low-HER2 head and neck cancer. In some embodiments, the cancer is low-HER2 breast cancer. In some embodiments, the cancer is low-HER2 gastric cancer. In some embodiments, the cancer is low-HER2 esophageal cancer. In some embodiments, the cancer is low-HER2 ovarian cancer. In some embodiments, the cancer is low-HER2 endometrial cancer. In some embodiments, the cancer is low-HER2 bladder cancer. In some embodiments, the cancer is low-HER2 lung cancer. In some embodiments, the cancer is low-HER2 cervical cancer. In some embodiments, the cancer is low-HER2 colon cancer. In some embodiments, the cancer is low-HER2 head and neck cancer.

[0249] In some implementations, the cancer is metastatic. In some implementations, the cancer is advanced. Example

[0250] Example 1. Combination of deuterated ETBR antagonist ICPI and CAR-T therapy in an in vivo model

[0251] Production of anti-HER2 mouse CAR-T cells

[0252] Lentiviral cells expressing anti-HER2 CAR were generated by co-transfecting 293T-17 cells with the lentiviral vector pLVX-EF1α-CAR-IRES-ZsGreen1 along with the packaging plasmids pSPAX2 and pMD2.G. Spleen cells from 6-week-old female BALB / c mice were separated by density gradient centrifugation using Histopaque lymphocyte separation medium and cultured in 24-well plates at 1×10⁻⁶ cells / well. 6 Cells were activated at a density of 10 cells / ml using a mouse T-cell activation / expansion kit for 2–3 days, and then transduced with PLVX-EF1α-CAR-IRES-ZsGreen1 lentivirus in the presence of 8 μg / mL polybrene, by centrifugation at 1,500 × g at 32 °C for 2 h at a multiplicity of infection of 20. GFP+ transduced T cells were analyzed by flow cytometry to assess lentiviral transduction efficiency.

[0253] To assess the cytotoxicity of anti-HER2 mouse CAR-T cells against HER2-positive breast cancer cells in vitro, CAR-T cells were incubated with 4T1-HER2 cells in triplicate at effector:target ratios of 2:1, 4:1, and 8:1. Specific lactate dehydrogenase (LDH) released from target cells into the culture supernatant was determined using an LDH cytotoxicity assay kit, according to the manufacturer's instructions. Additionally, IL-2 secretion from anti-HER2 mouse CAR-T cells was measured by incubating CAR-T cells with 4T1-HER2 cells in triplicate at E:T ratios of 2:1, 4:1, and 8:1 for 24 hours. IL-2 concentration in the culture supernatant was analyzed using an IL-2 ELISA kit (R&D Systems), according to the manufacturer's instructions.

[0254] 4T-1-HER2 mammary tumors were generated in syngeneic female BALB / c mice.

[0255] 4T1-HER2 cells (4×10) 6 Each tumor sample was resuspended in 0.1 mL of PBS and mixed with 0.1 mL of Matrigel, then subcutaneously injected into the right back region of female BALB / c mice (6 weeks old, weighing 17-20 g). Tumor growth was monitored every three days by measuring with calipers.

[0256] Treatment of 4T-1-HER2 breast tumors with anti-HER2 CAR-T and BQ-788-B

[0257] Approximately 14 days after injection of 4T-1-HER2 cells, when the tumor diameter reached approximately 6 mm, the mice were randomly divided into 8 groups for treatment (6 mice in each group).

[0258] Blank T or CAR-T cells (1×10⁻⁶)7 4T1-HER2 cells / mouse were administered intravenously to each mouse on days 11 and 18 post-tumor inoculation. Starting on day 10, BQ-788-B (24 μg) or PBS was administered subcutaneously to each mouse every 2 or 3 days for a total of 9 injections. From day 14 to day 34, all mice received an intraperitoneal injection of 20,000 IU IL-2 every 2 days (Note: Day 0 is defined as when 4T1-HER2 cells were injected into the mice). Starting on day 11 post-tumor inoculation, anti-PD1 or isotype control Ig (230 μg / mouse) was administered via IP injection every 4 days (for a total of 4 injections). Eight treatment groups are listed below:

[0259] 1) Blank T + PBS + Homotype

[0260] 2) CAR-T + PBS + Allogeneic

[0261] 3) BQ-788-B + Blank T + Same Type

[0262] 4) Blank T + PBS + Anti-PD1

[0263] 5) BQ-788-B+CAR-T+ Same type

[0264] 6) CAR-T + PBS + Anti-PD1

[0265] 7) BQ-788-B+ Blank T+ Anti-PD1

[0266] 8) BQ-788-B+CAR-T+Anti-PD1

[0267] Tumor growth was monitored every three days using calipers. On day 30 following injection of 4T1-HER2 cells, all mice were euthanized by CO2 asphyxiation followed by cervical dislocation. All tumors were dissected, photographed, and then fixed in 10% formalin.

[0268] result

[0269] Compared with the blank-T+PBS+isotype control group, all treatment groups showed a decrease in overall tumor growth ( Figure 2 On day 17, the combination of CAR-T, BQ-788-B, and anti-PD-1 therapy showed the greatest reduction in tumor growth, particularly compared to control or CAR-T alone. Figure 3A and Figure 3B At a later time point, BQ-788-B was added to enhance tumor suppression in mice treated with anti-PD1 or CAR-T therapy. On day 29 ( Figure 4Compared to the control group, CAR-T alone, or PD-1 alone, the group treated with BQ-788-B in combination with anti-PD-1 and / or CAR-T therapy had a significantly reduced tumor volume. On day 30 ( Figure 5A and Figure 5B Compared to the control group, CAR-T therapy alone, or PD-1 therapy alone, the group treated with BQ-788-B in combination with anti-PD-1 and / or CAR-T therapy had significantly reduced tumor weight. These experiments demonstrate that the addition of BQ-788-B significantly enhances the anti-tumor efficacy of CAR-T cells, anti-PD-1 antibody therapy, and the combination of CAR-T and anti-PD-1 antibody therapy.

[0270] Example 2. Treatment of solid tumors in human subjects with a combination therapy comprising CAR-expressing immune cells and a deuterated ETBR antagonist.

[0271] A combination therapy regimen comprising CAR-expressing immune cells (e.g., CAR-T or CAR-NK cells) and a deuterated ETBR antagonist (e.g., BQ-788-A, BQ-788-B, or BQ-788-C) is administered to human patients with cancer expressing a target (e.g., HER2-expressing cancer, such as breast cancer) according to the methods disclosed herein. This treatment reduces tumor volume or mass in the patient or eradicates the tumor.

[0272] Example 3. Treatment of solid tumors in human subjects with a combination therapy comprising CAR-expressing immune cells, immune checkpoint inhibitors, and deuterated ETBR antagonists.

[0273] A combination therapy regimen comprising CAR-expressing immune cells (e.g., CAR-T or CAR-NK cells), an ICPI (e.g., an anti-PD1 antibody), and a deuterated ETBR antagonist (e.g., BQ-788-A, BQ-788-B, or BQ-788-C) is administered to human patients with cancer (e.g., HER2-expressing cancer, such as breast cancer). This treatment reduces tumor volume or mass in the patient or eradicates the tumor in the patient.

[0274] While some embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Various changes, modifications, and substitutions will soon occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein can be used to practice the invention. The following claims are intended to define the scope of the invention and thereby cover the methods and structures within the scope of these claims and their equivalents.

Claims

1. A method of treating a cancer expressing a target in a subject with a corresponding need, the method comprising administering to the subject a combination comprising: (a) Immune cells expressing chimeric antigen receptors (CARs), wherein the CAR specifically binds to the target, and (b) An effective amount of ETBR antagonist.

2. In a method of treating a subject with cancer expressing a target using immune cells expressing a chimeric antigen receptor (CAR), wherein the CAR specifically binds to the target, the improvement includes co-administering an effective amount of an ETBR antagonist to the subject.

3. The method according to claim 1 or 2, wherein the target is a tumor-associated antigen.

4. The method according to claim 3, wherein the tumor-associated antigen is HER2.

5. The method according to any one of claims 1 to 4, further comprising administering an effective amount of an immune checkpoint inhibitor (ICPI) to the patient.

6. A method of treating a cancer expressing a target in a subject with a corresponding need, the method comprising administering to the subject a combination comprising: (a) Immune cells expressing chimeric antigen receptors (CARs), wherein the CARs specifically bind to the target. (b) Immune checkpoint inhibitors (ICPIs); and (c) An effective amount of ETBR antagonist.

7. In a method of treating a subject with a target cancer expressed by immune cells expressing a chimeric antigen receptor (CAR) and an immune checkpoint inhibitor (ICPI) in accordance with the need, wherein the CAR specifically binds to the target, the improvement includes co-administering an effective amount of an ETBR antagonist to the subject.

8. The method according to claim 6 or 7, wherein the target is a tumor-associated antigen.

9. The method of claim 8, wherein the tumor-associated antigen is HER2.

10. A method of treating HER2-expressing cancer in a subject with a corresponding need, the method comprising administering to the subject a combination comprising: (a) Immune cells expressing chimeric antigen receptor (CAR), wherein the CAR specifically binds to HER2, and (b) An effective amount of deuterated ETBR antagonist.

11. In a method of treating a subject with HER2-expressing cancer using immune cells expressing a chimeric antigen receptor (CAR), wherein the CAR specifically binds to HER2, the improvement includes co-administering an effective amount of an ETBR antagonist to the subject.

12. The method of claim 10 or 11 further comprises administering an effective amount of an immune checkpoint inhibitor (ICPI) to the patient.

13. The method according to any one of claims 5-9 and 12, wherein the ICPI is an anti-PD1 antibody, an anti-PD-L1 antibody, an anti-CTLA4 antibody, or a variant or functional fragment thereof.

14. The method of claim 13, wherein the ICPI is an anti-PD1 antibody.

15. The method according to any one of claims 1-14, wherein the ETBR antagonist is a deuterated ETBR antagonist.

16. The method of claim 15, wherein the deuterated ETBR antagonist is a compound of formula (1): Or its stereoisomers, wherein: n is an integer between 0 and 4; m is an integer between 0 and 3; X is a positively charged equilibrium ion; R1 and R3 are independently -H, -D, -CH3, -CH2D, -CHD2, or -CD3; R 2a R 2b R4, R5, and R6 are independently -CH3, -CH2D, -CHD2, or -CD3; and R1, R 2a R 2b Or at least one of R3 contains deuterium.

17. The method according to claim 16, wherein the compound is a compound of formula (1a): Or its stereoisomers.

18. The method according to claim 16, wherein the compound is a compound of formula (2): Or its stereoisomers.

19. The method according to claim 16, wherein the compound is a compound of formula (2a): Or its stereoisomers.

20. The method according to claim 16, wherein the compound is a compound of formula (3): Or its stereoisomers.

21. The method according to claim 16, wherein the compound is a compound of formula (3a): Or its stereoisomers.

22. The method according to claim 16, wherein the compound is a compound of formula (4): Or its stereoisomers.

23. The method according to claim 16, wherein the compound is a compound of formula (4a): Or its stereoisomers.

24. The method according to claim 16, wherein the compound is a compound of formula (5): Or its stereoisomers.

25. The method according to claim 16, wherein the compound is a compound of formula (5a): Or its stereoisomers.

26. The method according to claim 16, wherein the compound is a compound of formula (6): Or its stereoisomers.

27. The method according to claim 16, wherein the compound is a compound of formula (6a): Or its stereoisomers.

28. The method according to any one of claims 16, 18, 20, 22, 24, and 26, wherein n is 1, R1 is -D, and R 2a and R 2b Each is -CH3.

29. The method according to any one of claims 16-27, wherein R 2a It is -CH3, and R 2b It is -CH2D.

30. The method according to any one of claims 16-27, wherein R 2a and R 2b Each is -CH2D.

31. The method of claim 16, wherein n is 1, R1 is -D; and R 2a and R 2b Each is -CH2D.

32. The method according to any one of claims 1-31, wherein the deuterated ETBR antagonist is selected from: BQ-788-B BQ-788-A and BQ-788-C Or its stereoisomers.

33. The method according to any one of claims 1-32, wherein the ETBR antagonist is: BQ-788-B Or its stereoisomers.

34. The method according to any one of claims 1-33, wherein the ETBR antagonist is in a pharmaceutically acceptable excipient comprising dimethyl sulfoxide (DMSO).

35. The method according to any one of claims 1-34, wherein the CAR-expressing immune cell is a CAR-T cell or a CAR-NK cell.

36. The method of claim 35, wherein the CAR-expressing immune cell is a CAR-T cell.

37. The method according to any one of claims 1-36, wherein the cancer is an ETBR-associated cancer.

38. The method according to any one of claims 1-37, wherein the cancer is selected from the group consisting of: breast cancer, gastric cancer, esophageal cancer, ovarian cancer, endometrial cancer, bladder cancer, lung cancer, cervical cancer, colon cancer, and head and neck cancer.

39. The method according to any one of claims 1-9 and 13-32, wherein the cancer is a cancer expressing HER2.

40. The method of claim 39, wherein the cancer expressing HER2 is HER2+ cancer.

41. The method of claim 39, wherein the cancer expressing HER2 is a low-HER2 cancer.

42. The method according to any one of claims 39-41, wherein the cancer expressing HER2 is selected from the group consisting of: breast cancer, gastric cancer, esophageal cancer, ovarian cancer, endometrial cancer, bladder cancer, lung cancer, cervical cancer, colon cancer, and head and neck cancer.

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