Platelet storage methods and compositions

Abstract

Disclosed herein are compounds and compositions for slowing, preventing, or reversing platelet damage, particularly as may occur during blood banking or refrigerated storage of platelets. Also disclosed herein are methods for storing platelets and for improving platelet survival upon transfusion of one or more of the compounds or compositions described herein.

Description

[Technical field]

[0001] (Reference to Related Application) This patent application claims the benefit of priority to U.S. Provisional Application No. 62 / 961,580, filed January 15, 2020. All of the foregoing applications are fully incorporated by reference herein in their entirety for all purposes.

[0002] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT Financial support was provided in part by the following grants: NIH R01 HL147536, R43 HL123103, and UH54HL119810 (NCAI-CC).

[0003] (Technical field) This application relates to the fields of chemistry, biology, and medicine. Disclosed herein are chemical compositions and methods of use thereof. More specifically, disclosed herein are compounds, compositions for platelet storage, methods of platelet storage, and methods of using such stored platelets. [Background technology]

[0004] Platelet transfusion is a common life-saving procedure to treat or prevent bleeding in patients with low platelet counts or dysfunctional platelets, including cancer patients undergoing chemotherapy. Recently, the application of large doses of platelets during resuscitation has been shown to significantly improve the survival of trauma patients. In the United States alone, approximately 3 million doses of platelets are used annually. However, the current supply of platelets cannot effectively meet the increasing demand, mainly because, unlike other blood cells, platelets can only be stored at room temperature, but not refrigerated.

[0005] Room temperature storage favors bacterial growth, and the risk of bacterial infection for platelet concentrate transfusions is estimated to be 50 times higher than that for refrigerated red blood cell products. Thus, the US FDA limits room temperature storage of platelets to 7 days, with an actual shelf life of only 2.5-3 days, and a growing list of FDA-mandated tests for microbiological contamination after platelet collection from donors. Up to 20% of platelet preparations are discarded due to exhalation. However, a constant platelet shortage exists due to an unpredictable increase in use in surgical procedures, chemotherapy, and trauma situations. The short shelf life is a major obstacle to converting platelet products into a viable commodity, and although effective methods for safely storing human platelets for extended periods have been extensively studied, it remains a global unmet medical need for which there is no solution.

[0006] Frozen platelets (1-6oC) are hemostatically active but are rapidly cleared from the circulation after transfusion, and the mechanism of this clearance system has long been a mystery. The USCFR allows the use of cryogenic platelets for storage for up to 3 days for trauma patients (21 CFR 640.24 and 640.25).

[0007] The development of methods to prevent platelet damage during extended refrigeration is a critical need and has long been sought in blood banking. Such a development would revolutionize current methods of platelet storage. Summary of the Invention

[0008] Some embodiments provide a compound having a structure of formula (I): [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony: X 1 is optionally substituted aryl or optionally substituted heteroaryl; X 2 is selected from the group consisting of hydrogen, halogen, and optionally substituted alkyl; X3 is hydrogen or optionally substituted alkyl.

[0009] In some embodiments, X 1 is optionally substituted aryl. In some embodiments, X 1 is optionally substituted heteroaryl. In some embodiments, X 1 is substituted phenyl. In some embodiments, X 1 is 3,4-dichlorophenyl or 4-methylphenyl. 1 is unsubstituted heteroaryl. In some embodiments, X 1 is an unsubstituted 2-indole. In some embodiments, X 2 is hydrogen. In some embodiments, X 2 In some embodiments, X is chloride. 2 is trifluoromethyl. In some embodiments, X 3 is hydrogen. In some embodiments, X 3 is trifluoromethyl.

[0010] In some embodiments, the compound of Formula (I) is [ka] or a pharma- ceutically acceptable salt thereof.

[0011] Some embodiments relate to compositions for cold platelet storage at 0-20° C. In some embodiments, the composition comprises platelets, a compound of formula (I) or a pharma- ceutically acceptable salt thereof, or a combination thereof, and a physiologically acceptable carrier, the compound being present at a concentration of about 1 μM to about 20 μM. In some embodiments, the carrier is a buffer. In some embodiments, the carrier is selected from platelet additive solution (PAS), saline, phosphate buffered saline, Tris buffered saline, Hank's buffered saline, water, or a combination thereof. In some embodiments, the carrier comprises an electrolyte solution. In some embodiments, the composition comprises an additive selected from NaCl, KCl, CaCl2, MgCl2, MgSO4, NaCitrate, citric acid, NaHCO3, Na phosphate, Na acetate, Na gluconate, glucose, maltose, mannitol, and combinations thereof. In some embodiments, the additive is present in an amount of about 0.5 mmol / L to about 150 mmol / L. In some embodiments, the composition further comprises one or more components selected from D-ribose, D-glucose, Hank's solution, Hepes solution, bovine serum albumin, Tick anticoagulant peptide, and sterile water, or combinations thereof. In some embodiments, the composition further comprises an additive selected from a pH adjusting agent and a buffering agent, a tonicity agent, a stabilizer, a wetting agent, and combinations thereof. In some embodiments, the composition has a pH of about 5 to about 8. In some embodiments, the composition is isotonic.

[0012] Some embodiments relate to a method for storing platelets. In some embodiments, the method comprises storing the platelets in the presence of a compound of Formula (I) or a pharma- ceutically acceptable salt thereof. In some embodiments, the method comprises storing the platelets, comprising storing the platelets in a composition described herein. In some embodiments, the platelets are stored for 10 to 14 days. In some embodiments, the storage is performed at a temperature of about 0°C to about 10°C. In some embodiments, the storage is performed at a temperature of about 1°C to about 6°C. In some embodiments, the storage is performed at a temperature of about 2°C.

[0013] Some embodiments relate to a method for improving platelet survival during transfusion. In some embodiments, the method includes contacting platelets with a compound described herein or a pharma- ceutically acceptable salt thereof, or a composition described herein, or platelets stored according to a method described herein, and infusing the contacted platelets into a subject. In some embodiments, the compound is present at a concentration of about 1 μM to about 20 μM. In some embodiments, the platelets are stored in the presence of the compound at a temperature of about 1° C. to about 25° C. In some embodiments, the platelets are stored in the presence of the compound for about 1 to about 14 days. In some embodiments, the platelets are further contacted with a physiologically acceptable carrier. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] In one aspect, a compound is provided having the structure of formula (I): [ka]

[0015] In some embodiments, Formula (I) may be in the form of a pharma- ceutically acceptable salt as described herein.

[0016] In some embodiments of the compound of Formula (I), X 1 is selected from optionally substituted aryl, optionally substituted heteroaryl, substituted phenyl, 3,4-dichlorophenyl, 4-methylphenyl; X 1 is unsubstituted heteroaryl, or unsubstituted 2-indole. In some embodiments of the compounds of formula (I), X 1 is optionally substituted aryl. In some embodiments of the compounds of formula (I), X 1 is optionally substituted heteroaryl. In some embodiments of the compounds of formula (I), X 1 is a substituted phenyl. In some embodiments of the compounds of Formula (I), X 1is 3,4-dichlorophenyl or 4-methylphenyl. In some embodiments of the compound of formula (I), X 1 is unsubstituted heteroaryl. In some embodiments of the compounds of formula (I), X 1 is an unsubstituted 2-indole.

[0017] In some embodiments of the compound of Formula (I), X 2 is selected from hydrogen, deuterium, chloride, or trifluoromethyl. In some embodiments of the compounds of Formula (I), X 2 In some embodiments of the compound of formula (I), X 2 In some embodiments of the compounds of formula (I), X 2 is trifluoromethyl.

[0018] In some embodiments of the compound of Formula (I), X 3 is selected from hydrogen, deuterium, or trifluoromethyl. In some embodiments of the compounds of Formula (I), X 3 is hydrogen. In some embodiments of the compounds of formula (I), X 3 is trifluoromethyl.

[0019] In some embodiments, the compound of Formula (I) is [ka] or a pharma- ceutically acceptable salt thereof.

[0020] Some embodiments relate to compositions for cold platelet storage. In some embodiments, the compositions include platelets and a compound of formula (I) or a pharma- ceutically acceptable salt thereof. In some embodiments, the compositions include a combination of a compound of formula (I) and a pharma- ceutically acceptable salt thereof. In some embodiments, the compositions further include a carrier.

[0021] In some embodiments, storage is performed at a temperature greater than 0°C, 2°C, 4°C, 6°C, 8°C, 10°C, 12°C, 14°C, 16°C, 18°C, 20°C, 22°C, 24°C, 26°C, 28°C, 30°C, or at a range including and / or spanning the aforementioned values. In some embodiments, storage is performed at a temperature of about 0°C to about 25°C. In some embodiments, storage is performed at a temperature of about 0°C to about 20°C. In some embodiments, cold storage is performed at a temperature of about 0°C to about 10°C. In some embodiments, cold storage is performed at a temperature of about 1°C to about 6°C. In some embodiments, cold storage is performed at a temperature of about 2°C.

[0022] In some embodiments, the carrier is a physiologically acceptable carrier. In some embodiments, the carrier is a buffer. In some embodiments, the carrier is selected from platelet additive solution (PAS), saline, phosphate buffered saline, Tris buffered saline, Hank's buffered saline, water, or a combination thereof. In some embodiments, the carrier comprises an electrolyte solution. In some embodiments, the composition further comprises a pharma- ceutically acceptable excipient. In some embodiments, the composition further comprises a stabilizer. Such materials should be non-toxic and should not interfere with the efficacy of the active ingredient. Pharmaceutically acceptable excipients, carriers, buffers, or stabilizers can take a wide variety of forms, depending on the form of preparation desired for administration, e.g., intravenous or parenteral administration.

[0023] In some embodiments, the amount of carrier (in mg) is about 0.1, 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, or any amount including and / or ranging from the aforementioned values. In some embodiments, the amount of carrier present (in mg) is about 0.1, 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, or any amount including and / or ranging from the aforesaid values.

[0024] In some embodiments, the weight percentage of carrier in the composition is about 0, 1, 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, or more, or a range including and / or spanning the aforementioned values.

[0025] In some embodiments, the composition further comprises an additive. In some embodiments, the additive may be selected from pH adjusting and buffering agents, isotonicity agents, stabilizers, wetting agents, and combinations thereof. In some embodiments, the additive may be selected from NaCl, KCl, CaCl2, MgCl2, MgSO4, NaCitrate, citric acid, NaHCO3, Na phosphate, Na acetate, Na gluconate, glucose, maltose, mannitol, and combinations thereof. In some embodiments, the additive may be selected from D-ribose, D-glucose, Hank's solution, Hepes solution, bovine serum albumin, tick anticoagulant peptide, and sterile water, or combinations thereof.

[0026] In some embodiments, the additive is present in an amount including and / or exceeding the aforementioned values, including 0.5 mmol / L, 1.00 mmol / L, 5 mmol / L, 20 mmol / L, 35 mmol / L, 40 mmol / L, 55 mmol / L, 60 mmol / L, 70 mmol / L, 75 mmol / L, 85 mmol / L, 90 mmol / L, 95 mmol / L, 100 mmol / L, 110 mmol / L, 115 mmol / L, 125 mmol / L, 130 mmol / L, 140 mmol / L, 150 mmol / L, 165 mmol / L, 170 mmol / L, 175 mmol / L, 180 mmol / L, 185 mmol / L, 190 mmol / L, 195 mmol / L, 200 mmol / L, or any of the foregoing values. In some embodiments, the additive is present in an amount from about 0.5 mmol / L to about 150 mmol / L.

[0027] In some embodiments, the compound is administered at 0.1 μM, 1 μM, 2 μM, 3 μM, 4 μM, 8 μM, 9 μM, 12 μM, 15 μM, 16 μM, 20 μM, 24 μM, 25 μM, 26 μM, 28 μM, 30 μM, 34 μM, 35 μM, 36 μM, 38 μM, 40 μM, 42 μM, 45 μM, 46 μM, 50 μM, 52 μM, 55 μM, 56 μM, 58 μM, 5 μM, 76 μM, 80 μM, 82 μM, and / or ranges including the aforesaid values. In some embodiments, the compound is present in an amount of about 1 μM to about 100 μM, in some embodiments, the compound is present in an amount of about 1 μM to about 70 μM, in some embodiments, the compound is present in an amount of about 1 μM to about 20 μM, in some embodiments, the compound is present in an amount of about 1 μM to about 10 μM, and in some embodiments, the compound is present in an amount of about 1 μM to about 9 μM. In some embodiments, the compound is present in an amount of about 1 μM to about 5 μM. In some embodiments, the compound is present in an amount of about 5 μM to about 100 μM, in some embodiments, the compound is present in an amount of about 5 μM to about 70 μM, in some embodiments, the compound is present in an amount of about 5 μM to about 20 μM, in some embodiments, the compound is present in an amount of about 5 μM to about 10 μM, and in some embodiments, the compound is present in an amount of about 5 μM to about 9 μM. In some embodiments, the compound is present in an amount of about 10 μM to about 100 μM, in some embodiments, the compound is present in an amount of about 10 μM to about 70 μM, and in some embodiments, the compound is present in an amount of about 10 μM to about 20 μM.

[0028] In some embodiments, the composition has a pH of about 5, 5.5, 6.0, 6.5, 7.0, 7.5, 8, or a range including and / or spanning the aforementioned values. In some embodiments, the composition is isotonic.

[0029] Some embodiments relate to a method for storing platelets, in some embodiments, the method comprises storing said platelets in the presence of a compound of formula (I), in some embodiments, the method comprises storing said platelets in any of the aforementioned compositions.

[0030] In some embodiments, the platelets are stored for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, or a period of time ranging from and including the foregoing values. In some embodiments, the platelets are stored in the presence of the compound for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, or a period of time ranging from and including the foregoing values. In some embodiments, the platelets are stored in the presence of the compound for about 1 to about 14 days. In some embodiments, the platelets are stored in the presence of the compound for about 10 to about 14 days.

[0031] In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature greater than 0°C, 2°C, 4°C, 6°C, 8°C, 10°C, 12°C, 14°C, 16°C, 18°C, 20°C, 22°C, 24°C, 26°C, 28°C, 30°C, or in a range including and / or ranging from the aforementioned values. In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature of about 0°C to about 25°C. In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature of about 0°C to about 20°C. In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature of about 0°C to about 10°C. In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature of about 1°C to about 6°C. In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature of about 2°C.

[0032] Some embodiments relate to a method for improving platelet survival during transfusion, hi some embodiments, the method comprises contacting platelets with a compound of formula (I) or a pharma- ceutically acceptable salt thereof, and infusing the contacted platelets into a subject.

[0033] In some embodiments, the compound is present in an amount of 0.1 μM, 1 μM, 2 μM, 3 μM, 4 μM, 8 μM, 9 μM, 12 μM, 15 μM, 16 μM, 20 μM, 24 μM, 25 μM, 26 μM, 28 μM, 30 μM, 34 μM, 35 μM, 36 μM, 38 μM, 40 μM, 42 μM, 45 μM, 46 μM, 50 μM, 52 μM, 55 μM, 56 μM, 58 μM, 60 μM, 62 μM, 65 μM, 66 μM, 68 μM, 70 μM, 74 μM or more. 5 μM, 76 μM, 80 μM, 82 μM, 84 μM, 85 μM, 86 μM, 88 μM, 90 μM, 92 μM, 94 μM, 95 μM, 96 μM, 98 μM, 100 μM, 105 μM, 110 μM, 115 μM, 120 μM, 125 μM, 130 μM, 140 μM, 150 μM, 160 μM, 170 μM, 180 μM, 190 μM, 200 μM, or ranges including and / or spanning the aforesaid values. In some embodiments, the compound is present in an amount of about 1 μM to about 100 μM, in some embodiments, the compound is present in an amount of about 1 μM to about 70 μM, in some embodiments, the compound is present in an amount of about 1 μM to about 20 μM, in some embodiments, the compound is present in an amount of about 1 μM to about 10 μM, and in some embodiments, the compound is present in an amount of about 1 μM to about 9 μM. In some embodiments, the compound is present in an amount of about 1 μM to about 5 μM. In some embodiments, the compound is present in an amount of about 5 μM to about 100 μM, in some embodiments, the compound is present in an amount of about 5 μM to about 70 μM, in some embodiments, the compound is present in an amount of about 5 μM to about 20 μM, in some embodiments, the compound is present in an amount of about 5 μM to about 10 μM, and in some embodiments, the compound is present in an amount of about 5 μM to about 9 μM. In some embodiments, the compound is present in an amount of about 10 μM to about 100 μM, in some embodiments, the compound is present in an amount of about 10 μM to about 70 μM, and in some embodiments, the compound is present in an amount of about 10 μM to about 20 μM.

[0034] In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature greater than 0°C, 2°C, 4°C, 6°C, 8°C, 10°C, 12°C, 14°C, 16°C, 18°C, 20°C, 22°C, 24°C, 26°C, 28°C, 30°C, or in a range including and / or ranging from the aforementioned values. In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature of about 0°C to about 25°C. In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature of about 0°C to about 20°C. In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature of about 0°C to about 10°C. In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature of about 1°C to about 6°C. In some embodiments, the platelets are stored in the presence of a compound of Formula (I) at a temperature of about 2°C.

[0035] In some embodiments, the platelets are stored for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, or a period of time ranging from and including the foregoing values. In some embodiments, the platelets are stored in the presence of the compound for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, or a period of time ranging from and including the foregoing values. In some embodiments, the platelets are stored in the presence of the compound for about 1 to about 14 days. In some embodiments, the platelets are stored in the presence of the compound for about 10 to about 14 days.

[0036] In some embodiments, the platelets are further contacted with a physiologically acceptable carrier.

[0037] Some embodiments relate to compositions comprising a compound of Formula (I) and a platelet additive solution (PAS), wherein each 100 mL of PAS contains 0.405 g sodium chloride, 0.037 g potassium chloride, 0.030 g magnesium chloride 6H2O, 0.442 g acetate 3H2O, 0.318 g sodium citrate 2H2O, 0.093 g sodium phosphate dihydrogen 1H2O, 0.769 g sodium phosphate disodium 12H2O, and 100 mL water for injection.

[0038] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the embodiments belong. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the embodiments, the preferred methods and materials are described here. All publications mentioned herein are expressly incorporated by reference in their entirety.

[0039] When the compounds disclosed herein have at least one chiral center, they can exist as individual enantiomers and diastereomers, or as mixtures of such isomers, including racemates. Separation of individual isomers or selective synthesis of individual isomers can be achieved by application of various methods well known to those skilled in the art. Unless otherwise specified, all such isomers and mixtures thereof are included within the scope of the compounds disclosed herein. Furthermore, the compounds disclosed herein can exist in one or more crystalline or amorphous forms. Unless otherwise specified, all such forms are included within the scope of the compounds disclosed herein, including any polymorphic forms. Furthermore, some of the compounds disclosed herein can form solvates with water (i.e., hydrates) or common organic solvents. Unless otherwise specified, such solvates are included within the scope of the compounds disclosed herein.

[0040] Those skilled in the art will recognize that some of the structures depicted herein may be resonance forms or tautomers of compounds that may be significantly, even kinetically, represented by other chemical structures; those skilled in the art will recognize that such structures may represent only a very small portion of a sample of such compounds. Although such compounds are considered to be within the scope of the depicted structures, such resonance forms or tautomers are not represented herein.

[0041] Isotopes may be present in the compounds described. Each chemical element as represented in a compound structure may include any isotope of said element. For example, in a compound structure, a hydrogen atom may be explicitly disclosed or understood to be present in the compound. At any position of the compound where a hydrogen atom may be present, the hydrogen atom may be any isotope of hydrogen, including but not limited to hydrogen-1 (protium) and hydrogen-2 (deuterium). Thus, reference to a compound herein encompasses all possible isotopic forms unless the context clearly dictates otherwise.

[0042] definition As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, a reference to "platelets" includes a plurality of such platelets, a reference to "a carrier" includes a reference to one or more carriers and equivalents thereof known to those skilled in the art, and so forth.

[0043] "Solvate" refers to a compound formed by the interaction of a solvent with a compound described herein, or a metabolite, or salt thereof. Suitable solvates are pharma- ceutically acceptable solvates, including hydrates.

[0044] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness and properties of a compound, and is not undesirable for use pharmaceutical or otherwise. In many cases, the compounds herein can form acid salts and / or base salts due to the presence of amino and / or carboxyl groups or groups similar thereto. Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc., with ammonium, potassium, sodium, calcium and magnesium salts being particularly preferred. Organic bases from which salts can be derived include, for example, primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, etc., specifically isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine and ethanolamine. Many such salts are known in the art, as described in International Publication WO 87 / 05297, published September 11, 1987 by Johnston et al., which is incorporated herein by reference in its entirety.

[0045] As used herein, "C" refers to a set of integers where "a" and "b" are integers. b " or "C a-b " to "C a " refers to the number of C atoms in a particular group. That is, a group can contain from "a" to "b" carbon atoms (inclusive). Thus, for example, "C1-C4 alkyl" or "C1-4 An "alkyl" group refers to all alkyl groups having 1 to 4 carbons, i.e., CH3-, CH3CH2-, CH3CH2CH2-, (CH3)2CH-, CH3CH2CH2CH2-, CH3CH2CH(CH3)-, and (CH3)3C-.

[0046] The terms "halogen" or "halo" as used herein refer to any one of the radiostable atoms in column 7 of the periodic table of the elements, e.g., fluorine, chlorine, bromine, or iodine, with fluorine and chlorine being preferred.

[0047] As used herein, "alkyl" refers to a straight or branched hydrocarbon chain that is fully saturated (i.e., contains no double or triple bonds). The alkyl group may have 1 to 20 carbon atoms (wherever it appears herein, a numerical range such as "1 to 20" refers to each integer within the given range, e.g., "1 to 20 carbon atoms" means that the alkyl group consists of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms, although the definition specifies numerical ranges). The alkyl group may also be a medium-sized alkyl having 1 to 9 carbon atoms. The alkyl group may be a lower alkyl having 1 to 4 carbon atoms. The alkyl group of the compound may be a "C 1-4 By way of example only, "C 1-4 "Alkyl" indicates that there are 1 to 4 carbon atoms in the alkyl chain, i.e., the alkyl chain is selected from the group consisting of methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, and t-butyl. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl, and the like.

[0048] As used herein, "alkoxy" refers to the formula -OR, where R is alkyl as defined above, e.g., "C 1-9Alkoxy" includes, but is not limited to, methoxy, ethoxy, n-propoxy, 1-methylethoxy (isopropoxy), n-butoxy, iso-butoxy, sec-butoxy, and tert-butoxy.

[0049] As used herein, "heteroalkyl" refers to a straight or branched hydrocarbon chain containing one or more heteroatoms, i.e., elements other than carbon, including, but not limited to, nitrogen, oxygen, and sulfur, in the chain backbone. Heteroalkyl groups can have from 1 to 20 carbon atoms, although the definition also encompasses the occurrence of the term "heteroalkyl" where no numerical range is specified. Heteroalkyl groups can also be medium size heteroalkyls having from 1 to 9 carbon atoms. Heteroalkyl groups can also be lower heteroalkyls having from 1 to 4 carbon atoms. The heteroalkyl groups of a compound are defined as "C 1-4 A heteroalkyl group may contain one or more heteroatoms. 1-4 "Heteroalkyl" indicates that there are 1 to 4 carbon atoms in the heteroalkyl chain and further that there are one or more heteroatoms in the backbone of the chain.

[0050] The term "aromatic" refers to a ring or ring system having a conjugated pi-electron system, and includes both carbocyclic aromatic (e.g., phenyl) and heterocyclic aromatic (e.g., pyridine) groups. The term includes monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of atoms) groups, so long as the entire ring system is aromatic.

[0051] As used herein, "aryl" refers to an aromatic ring or ring system (i.e., two or more fused rings that share two adjacent carbon atoms) that contains only carbon in the ring backbone. When aryl is a ring system, all rings in the system are aromatic. An aryl group can have from 6 to 18 carbon atoms, although the definition also encompasses occurrences of the term "aryl" where no numerical range is specified. In some embodiments, an aryl group has from 6 to 10 carbon atoms. An aryl group is defined as "C 6-10Aryl, C6 or C 10 Examples of aryl groups include, but are not limited to, phenyl, naphthyl, azulenyl, and anthracenyl.

[0052] As used herein, "aryloxy" and "arylthio" refer to RO- and RS-, where R is aryl as defined above, e.g., "C 6-10 aryloxy" or "C 6-10 and "arylthio", including, but not limited to, phenyloxy.

[0053] "Aralkyl" or "arylalkyl" refers to an aryl group linked via an alkylene group as a substituent, e.g., "C 7-14 In some cases, the alkylene group may be a lower alkylene group (i.e., C 1-4 alkylene group).

[0054] As used herein, "heteroaryl" refers to an aromatic ring or ring system (i.e., two or more fused rings sharing two or more adjacent atoms) containing one or more heteroatoms, i.e., elements other than carbon, including, but not limited to, nitrogen, oxygen, and sulfur, in the ring backbone. When heteroaryl is a ring system, all rings in the system are aromatic. Heteroaryl groups can have 5 to 18 ring members (i.e., the number of atoms that make up the ring backbone, including carbon atoms and heteroatoms), although this definition also encompasses the existence of the term "heteroaryl" where no numerical range is specified. In some embodiments, heteroaryl groups have 5 to 10 ring members or 5 to 7 ring members. Heteroaryl groups can be referred to as "5-7 membered heteroaryl", "5-10 membered heteroaryl", or similar designations. Examples of heteroaryl rings include, but are not limited to, furyl, thienyl, phthalazinyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, triazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, quinolinyl, isoquinylinyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, indolyl, isoindolyl, and benzothienyl.

[0055] A "heteroaralkyl" or "heteroarylalkyl" is a heteroaryl group bonded as a substituent via an alkylene group. Examples include, but are not limited to, 2-thienylmethyl, 3-thienylmethyl, furylmethyl, thienylethyl, pyrrolylalkyl, pyridylalkyl, isoxazolylalkyl, and imidazolylalkyl. In some cases, the alkylene group may be a lower alkylene group (i.e., C 1-4 alkylene group).

[0056] As used herein, "carbocyclyl" refers to a non-aromatic ring or ring system that contains only carbon atoms in the ring system backbone. When carbocyclyl is a ring system, two or more rings can be linked together in a fused, bridged, or spiro-linked fashion. Carbocyclyl can have any degree of saturation, provided that at least one ring in the ring system is not aromatic. Thus, carbocyclyl includes cycloalkyl, cycloalkenyl, and cycloalkynyl. Carbocyclyl groups can have 3 to 20 carbon atoms, although this definition also encompasses the existence of the term "carbocyclyl" where no numerical range is specified. Carbocyclyl groups can also be medium-sized carbocyclyls having 3 to 10 carbon atoms. Carbosyl groups can also be carbocyclyls having 3 to 6 carbon atoms. A carbocyclyl group is a "C 3-6 ·carbocyclyl" or similar names. Examples of carbocyclyl rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, 2,3-dihydro-indene, bicyclo[2.2.2]octanyl, adamantyl, and spiro[4.4]nonanyl.

[0057] "(Carbocyclyl)alkyl" refers to a carbocyclyl group bonded as a substituent via an alkylene group, e.g., "C 4-10 (carbocyclyl)alkyl" including, but not limited to, cyclopropylmethyl, cyclobutylmethyl, cyclopropylethyl, cyclopropylbutyl, cyclobutylethyl, cyclopropylisopropyl, cyclopentylmethyl, cyclopentylethyl, cyclohexylmethyl, cyclohexylethyl, cycloheptylmethyl, and the like. In some instances, the alkylene group is a lower alkylene group.

[0058] As used herein, "cycloalkyl" means a fully saturated carbocyclyl ring or ring system. Examples include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0059] As used herein, "cycloalkenyl" refers to a carbocyclyl ring or ring system that has at least one double bond, and no ring in the ring system is aromatic, for example, cyclohexenyl.

[0060] As used herein, "heterocyclyl" refers to a non-aromatic cyclic ring or ring system containing at least one heteroatom in the ring backbone. Heterocyclyls may be joined together in a fused, bridged, or spiro-linked fashion. Heterocyclyls may have any degree of saturation, provided that at least one ring in the ring system is not aromatic. The heteroatom may be present in either the non-aromatic or aromatic ring in the ring system. Heterocyclyl groups may have 3-20 ring members (i.e., the number of atoms that make up the ring backbone, including carbon atoms and heteroatoms), although this definition also encompasses the existence of the term "heterocyclyl" where no numerical range is specified. Heterocyclyl groups may also be medium-sized heterocyclyls having 3-10 ring members. Heterocyclyl groups may also be heterocyclyls having 3-6 ring members. Heterocyclyl groups may be designated as "3-6 membered heterocyclyl" or similar names. In preferred 6-membered monocyclic heterocyclyls the heteroatom is selected from one of O, N or S, and in preferred 5-membered monocyclic heterocyclyls the heteroatom is selected from one or two of O, N or S. Examples of heterocyclyl rings are azepinyl, acridinyl, carbazolyl, cinnolinyl, dioxolanyl, imidazolinyl, imidazolidinyl, morpholinyl, oxiranyl, oxypanyl, thiepanyl, piperidinyl, piperazinyl, dioxopiperazinyl, pyrrolidinyl, pyrrolidionyl, 4-piperidonyl, pyrazolinyl, pyrazolidinyl, 1,3-dioxinyl, 1,4-dioxinyl, 1,3-oxathianyl, 1,4-oxathianyl, and 2H-1,2-oxazinyl. Examples include, but are not limited to, trioxanyl, hexahydro-1,3,5-triazinyl, 1,3-dioxolyl, 1,3-dithiolanyl, 1,3-dithiolanyl, isoxazolinyl, isoxazolidinyl, oxazolinyl, oxazolidinonyl, thiazolinyl, thiazolidinonyl, 1,3-oxathiolanyl indolinyl, isoindolinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydro-1,4-thiazinyl, thiamorpholinyl, dihydrobenzofuranyl, benzimidazolidinyl, and tetrahydroquinoline.

[0061] "(Heterocyclyl)alkyl" is a heterocyclyl group attached as a substituent via an alkylene group. Examples include, but are not limited to, imidazolinylmethyl and indolinylethyl.

[0062] As used herein, "acyl" refers to -C(=O)R, where R is hydrogen, C, 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 carbocyclyl, aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl. Non-limiting examples include formyl, acetyl, propanoyl, benzoyl, and acryl.

[0063] An "O-carboxy" group refers to a "-OC(=O)R" group, where R is hydrogen, C, 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 It is selected from carbocyclyl, aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0064] A "C-carboxy" group refers to a "-C(=O)OR" group, where R is hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 It is selected from carbocyclyl, aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl. A non-limiting example is carboxyl (i.e., -C(=O)OH).

[0065] A "cyano" group refers to a "-CN" group.

[0066] A "cyanato" group refers to a "-OCN" group.

[0067] An "isocyanato" group refers to a "-NCO" group.

[0068] A "thiocyanato" group refers to a "-SCN" group.

[0069] An "isothiocyanato" group refers to a "-NCS" group.

[0070] A "sulfinyl" group refers to a "-S(=O)R" group, where R is hydrogen, C, 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclyl, C 6-10 It is selected from aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0071] A "sulfonyl" group refers to a "-SO2R" group, where R is hydrogen, C, 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclyl, C 6-10 It is selected from aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0072] The "S-sulfonamide" group is "-SONR A R B " group, where R A and R B are each independently hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclyl, C 6-10 It is selected from aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0073] An “N-sulfonamide” group is defined as “-N(R A )SO2R B " group, where R A and Rb are each independently, as defined herein, hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclyl, C 6-10 It is selected from aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0074] The "O-carbamyl" group is "-OC(=O)NR A R B " group, where R A and R B are each independently hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclyl, C 6-10 It is selected from aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0075] The "N-carbamyl" group is defined as "-N(R A )OC(=O)R B " group, where R A and R B are each independently hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclyl, C 6-10 It is selected from aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0076] The "O-thiocarbamyl" group is "-OC(=S)NR A R B " group, where R A and R B are each independently hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclyl, C6-10 It is selected from aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0077] An "N-thiocarbamyl" group refers to a "-N(RA)OC(=S)RB" group, where RA and RB are each independently hydrogen, C 1-6 Alkyl, C 2-6 alkenyl.

[0078] C 2-6 Alkynyl, C 3-7 Carbocyclyl, C 6-10 Aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl are as defined herein.

[0079] A "C-amide" group is -C(=O)NR A R B " group, where R A and R B are each independently hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclyl, C 6-10 It is selected from aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0080] An "N-amide" group is an -N(R A )C(=O)R B " group, where R A and R B are each independently hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclyl, C 6-10 It is selected from aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0081] The "amino" group is "-NR A R B" group, where R A and R B are each independently hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclyl, C 6-10 It is selected from aryl, 5- to 10-membered heteroaryl, and 5- to 10-membered heterocyclyl.

[0082] An "aminoalkyl" group refers to an amino group bonded via an alkylene group.

[0083] An "alkoxyalkyl" group refers to an alkoxy group linked via an alkylene group, e.g., "C 2-8 "Alkoxyalkyl" and the like.

[0084] As used herein, a substituent is derived from an unsubstituted parent group in which one or more hydrogen atoms have been replaced with another atom or group. Unless otherwise indicated, when a group is considered to be "substituted", C1-C6 alkyl, C1-C6 alkyl, C1-C6 alkynyl, C1-C6 heteroalkyl, haloalkyl, C3-C7 carbocyclyl (optionally substituted with halo, C1-C6 alkyl, C1-C6 haloalkyl, and C1-C6 haloalkoxy), C3-C7-carbocyclyl-C1-C6-alkyl (optionally substituted with halo, C1-C6 alkyl, C1-C6 alkoxy, and C1-C6 haloalkoxy), 5-10 membered heterocyclyl (halo, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy) , C1-C6 alkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy), 5-10 membered heterocyclyl-C1-C6-alkyl, aryl (optionally substituted with halo, C1-C6 alkoxy, C1-C6 haloalkoxy, and C3-C7 haloalkoxy), (halo, alkylalkoxy, C1-C6 haloalkyl, and C1-C6 haloalkoxy), 5-10 membered heteroaryl arylC1-C6 alkyl (optionally substituted with halo), 5-10 membered heteroaryl (optionally substituted with halo, C1-C6 alkoxy, C1-C6 haloalkoxy, and C3-C7 haloalkoxy), 5-10 membered heteroarylC1-C6 alkyl (optionally substituted with C1-C6 alkoxy, C1-C6 haloalkoxy, and C3-C7 haloalkoxy), halo, cyano, hydroxy, C1-C6 alkoxy, C1-C6 alkoxy(C1-C6)alkyl (i.e., ether), aryloxy (optionally substituted with C1-C6 alkoxy, C1-C6 haloalkoxy, and C3-C7 haloalkoxy), sulfhydryl (mercapto), halo(C1-C6)alkyl (e.g., -CF3), halo(C1-C6)alkoxy (e.g., -OCF3), C1-C6 alkylthio, arylthio, amino, amino(C1-C6)alkyl, nitro, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido. Examples include S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, acyl, cyanato, isocyanato, thiocyanato, isothiocyanato, sulfinyl, sulfonyl, and oxo (=O). When a group is described as "optionally substituted," it means that the group is optionally substituted with those substituents.

[0085] In some embodiments, the substituents are substituted with one or more substituents individually and independently selected from C1-C4 alkyl, amino, hydroxy, and halogen.

[0086] It is understood that certain radical naming conventions can include either mono-radicals or di-radicals, depending on the context. For example, if a substituent requires two points of attachment to the remainder of the molecule, the substituent is understood to be a diradical. For example, a substituent identified as an alkyl, which requires two points of attachment, includes diradicals such as -CH2-, -CH2CH2-, -CH2CH(CH3)CH2-, etc. Other radical naming conventions explicitly indicate that the radical is a diradical, such as "alkylene" or "alkenylene."

[0087] Whenever a substituent is depicted as a diradical (i.e., having two points of attachment to the remainder of the molecule), it is understood that the substituent may be attached in any orientation unless otherwise indicated. Thus, for example, a substituent depicted as -AE-, or [ka] Includes substituents where A is oriented such that it is attached to the leftmost attachment point of the molecule, as well as where A is attached to the rightmost attachment point of the molecule.

[0088] The term "platelets" is used herein to refer to blood platelets. Platelets can be described as miniature plasma disks present in the blood of vertebrates. Platelets play a role in blood clotting. Platelets can be derived from any source, including the human blood supply or the patient's own blood.

[0089] When a range of values ​​is provided, it is understood that the upper and lower limits, as well as each intervening value between the upper and lower limits of the range, are included within an embodiment.

[0090] The term "about" or "approximately" means within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, "about" can mean within 1 or more than 1 standard deviation, according to the practice in the art. Alternatively, "about" can mean within a range of up to 20%, up to 10%, up to 5%, and up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean values ​​within an order of magnitude, within 5-fold, and within 2-fold. When a particular value is described in the present application and claims, unless otherwise indicated, the term "about" should be assumed to mean within an acceptable error range for that particular value.

[0091] As used herein, the term "effective amount" refers to an amount of one or more active ingredients sufficient to produce a desired effect. This includes both therapeutic and prophylactic effects. When referring to an individual active ingredient administered alone, the term refers to that ingredient alone. When referring to a combination, the term refers to the combined amount of active ingredients that produces a therapeutic effect, regardless of whether the active ingredients are administered in combination, sequentially, or simultaneously.

[0092] It is understood that the methods and formulations described herein include the use of pharma- ceutically acceptable salts and / or conformers of the compounds of the disclosed embodiments, as well as metabolites and active metabolites of these compounds that have the same type of activity. Conformers are structures that are conformers. Conformation is the phenomenon of molecules that have the same structural formula but have different conformations (conformers) of atoms around a rotating bond. Similarly, it is understood that the compounds described herein include any form of the compounds described herein (e.g., pharma- ceutically acceptable salts, enantiomeric forms, tautomeric forms, etc.).

[0093] The term "pharmaceutical composition" refers to a mixture of one or more compounds disclosed herein with other chemical components, such as diluents or carriers. A pharmaceutical composition facilitates administration of a compound to an organism. A pharmaceutical composition is generally tailored for a specific intended route of administration.

[0094] The term "pharmaceutical acceptable salt" refers to a salt of a compound that does not cause significant irritation to an organism to which it is administered and does not abrogate the biological activity and properties of the compound. In some embodiments, the salt is an acid addition salt of the compound. Pharmaceutical salts can be obtained by reacting the compound with inorganic acids such as hydrohalic acids (e.g., hydrochloric acid or hydrobromic acid), sulfuric acid, nitric acid, and phosphoric acid. Pharmaceutical salts can also be obtained by reacting the compound with organic acids, such as aliphatic or aromatic carboxylic or sulfonic acids, such as formic acid, acetic acid, succinic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, nicotinic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, or naphthalenesulfonic acid. Pharmaceutical salts can also be obtained by reacting the compounds with bases to form ammonium salts, alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as calcium or magnesium salts, salts of organic bases such as dicyclohexylamine, N-methyl-D-glucamine, Tris(hydroxymethyl)methylamine, C1-C7 alkylamines, cyclohexylamine, triethanolamine, ethylenediamine, and salts with amino acids such as arginine and lysine.

[0095] As used herein, "carrier" refers to a compound that facilitates the delivery of a compound to a cell or tissue. For example, but not limited to, dimethyl sulfoxide (DMSO) is a commonly used carrier that facilitates the uptake of many organic compounds into cells or tissues of a subject.

[0096] As used herein, "excipient" refers to an inert substance added to a pharmaceutical composition to provide the composition with bulk, consistency, stability, binding ability, lubricity, disintegration ability, etc. A "diluent" is a type of excipient.

[0097] As used herein, the term "weight percent" when referring to a component is the weight of the component divided by the weight of the composition containing the component multiplied by 100%. For example, when 5 grams of component A is added to 95 grams of component B, the weight percent of component A is 5% (e.g., 5g A / (5g A+95g B)×100%).

[0098] The mechanisms of how low temperature affects platelet survival are not fully understood. The effects of low temperature on platelets are complex and are believed to include shape changes, cytoskeletal reorganization, activation, cell surface protein clustering, and changes in the carbohydrate structure of surface glycoproteins. Cold storage is believed to shorten platelet lifespan due to a decrease in temperature that clusters glycoprotein-Ib (GPIb) receptors on specific microdomains of the platelet membrane. Applicants have found that recognition of specific glycated / sialylated residues on clustered glycoproteins by macrophage β2 integrins and hepatocyte Ashwell-Morell receptors leads to platelet phagocytosis by the host and removal from circulation. Applicants hypothesize that changes in Rho GTPase activity may affect platelet membrane lipid raft assembly and glycoprotein composition. Reversible targeting of Rho family GTPases with small molecule inhibitors may prevent cytoskeleton-dependent cold storage pathology in platelets and result in increased platelet survival.

[0099] Disclosed herein are compositions and methods useful for platelet survival and / or quality, transfusion, and related issues. In one aspect, a composition for platelet storage or treatment is described.

[0100] The active agents described herein can form salts which are also within the scope of the preferred embodiments, and references herein to the compounds of the active agents are understood to include references to the salts thereof, unless otherwise indicated. EXAMPLES

[0101] Synthesis of UC-177633 UC-177633 was synthesized according to the following scheme. [ka]

[0102] UC-177633 was synthesized in two steps:

[0103] Step 1: 4-Chloro-8-(trifluoromethyl)quinolone was reacted with 5 mol% PD[P(o-tolyl)3]2, CyPF-tBu, potassium hydroxylase, and 2 equivalents of hydrazine in dioxane at 100 °C for 8 h to produce 4-hydrazine-8-(trifluoromethyl)quinoline.

[0104] Step 2: The obtained 4-hydrazinyl-8-(trifluoromethyl)quinolone was reacted with an equal amount of 4-methylbenzoic acid, 0.1 mmol of HATU (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate), and DMR at 0° C. for 1 hour and allowed to stand at room temperature overnight. The reaction mixture was diluted with ethyl acetate, and the mixture was washed successively with 5% aqueous citric acid, saturated sodium bicarbonate solution, and saturated sodium chloride solution. The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was removed under vacuum. The crude product was crystallized to give UC-177633.

[0105] Additional compounds listed in Table 1 are synthesized following a similar sequence of steps, with structural variations provided by utilizing quinoline starting materials or aromatic / heteroaromatic compound variants added at the coupling step. Those skilled in the art will recognize that similar compounds can be synthesized using similar synthetic schemes. Those skilled in the art will recognize that the compounds of this embodiment can be synthesized using other synthetic schemes.

[0106] Synthesis of UC-177617 UC-177617 was synthesized according to the following scheme. [ka]

[0107] 4-Iodoquinoline was added to 1.5 equivalents of 3,4-dichlorobenzohydrazide. The reaction mixture was subjected to CuI-catalyzed reaction with 0.1 equivalents of CuI, 0.2 equivalents of potassium carbonate in DMSO at 60 °C for 20-30 h. The resulting product gave rise to UC-177617 with an efficiency of about 70%.

[0108] Additional compounds listed in Table 1 may be synthesized following a similar sequence of steps, if there are structural variations.Those skilled in the art will recognize that similar compounds may be synthesized using similar synthetic schemes.Those skilled in the art will recognize that the compounds of this embodiment may be synthesized using other synthetic schemes.

[0109] Inhibition of collagen-induced platelet aggregation Platelet shape changes of washed platelets were monitored using an aggregometer and the decrease in light transmittance after addition of agonists (Huzoor et al., 1993). For the purpose of this analysis, collagen was used as the agonist. Briefly, washed human platelet suspensions were incubated at 37°C with stirring at 900 rpm in a Lumi-Aggregometer (Chrono-Log Corporation), followed by the addition of dimethyl sulfoxide (DMSO) as a control vehicle, or various compounds dissolved in DMSO. After incubation of the samples with the respective compounds for 2 min, platelet aggregation was induced by the addition of collagen (1-4 μg / ml) and relative aggregation values ​​were read 6 min after induction. A dose response for each compound was derived. These data were used to determine the IC at which 50% aggregation was achieved. 50 The concentrations were determined. The results of these experiments are shown in Table 1. [Table 1] JPEG0007628714000009.jpg62170

Claims

1. Compounds of formula (I): 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof, During the ceremony: X １ is selected from 3,4-dichlorophenyl, 4-methylphenyl, or unsubstituted 2-indole; X ２ is selected from the group consisting of hydrogen, deuterium, halogen, and optionally substituted alkyl; X ３ is hydrogen, deuterium, or optionally substituted alkyl.

2. X １ The compound according to claim 1, wherein is 3,4-dichlorophenyl or 4-methylphenyl.

3. X １ The compound of claim 1, wherein is an unsubstituted 2-indole.

4. X ２ The compound according to any one of claims 1 to 3, wherein is hydrogen or deuterium.

5. X ２ The compound according to any one of claims 1 to 3, wherein is chloride.

6. X ２ The compound according to any one of claims 1 to 3, wherein is trifluoromethyl.

7. X ３ The compound according to any one of claims 1 to 6, wherein is hydrogen.

8. X ３ The compound according to any one of claims 1 to 6, wherein is trifluoromethyl.

9. 【Chemical 2】 or a pharma- ceutically acceptable salt thereof.

10. 1. A composition for cold platelet storage, comprising: platelet; A compound according to any one of claims 1 to 9, or a pharma- ceutically acceptable salt thereof, or a combination thereof; and Physiologically acceptable carriers.

11. The composition of claim 10, wherein the cold platelet storage composition is at a temperature of 0-20°C.

12. The composition according to claim 10 or claim 11, wherein the storage is at a temperature between 0°C and 10°C.

13. The composition according to claim 10 or claim 11, wherein the storage is at a temperature between 1°C and 6°C.

14. 12. The composition of claim 10 or claim 11, wherein the storage is at a temperature of 2°C.

15. The composition of any one of claims 10 to 14, wherein the compound is present in an amount of 1 μM to 20 μM.

16. The composition of any one of claims 10 to 15, wherein the compound is present in an amount of 1 μM to 9 μM.

17. The composition of any one of claims 10 to 16, wherein the compound is present in an amount of 1 μM to 5 μM.

18. The composition of any one of claims 10 to 17, further comprising an additive.

19. The additives are NaCl, KCl, CaCl ２ , MgCl ２ , MgSO ４ , Sodium Citrate ３ , Citric Acid, NaHCO ３ 20. The composition of claim 18, wherein the sugar is selected from the group consisting of sodium phosphate, sodium acetate, sodium gluconate, glucose, maltose, mannitol, and combinations thereof.

20. The composition of any one of claims 10 to 19, wherein the physiologically acceptable carrier is a buffering agent.

21. 20. The composition of any one of claims 10 to 19, wherein the physiologically acceptable carrier is selected from platelet additive solution (PAS), saline, phosphate buffered saline, Tris buffered saline, Hank's buffered saline, water, or a combination thereof.

22. The composition of any one of claims 10 to 19, wherein the physiologically acceptable carrier comprises an electrolyte solution.

23. The composition of any one of claims 18 to 22, further comprising an additive selected from pH adjusting and buffering agents, tonicity adjusting agents, stabilizers, wetting agents, and combinations thereof.

24. The composition according to any one of claims 10 to 23, wherein the composition has a pH of from 5 to 8.

25. The composition according to any one of claims 10 to 14, wherein the composition is isotonic.

26. 10. A composition for cold platelet storage at 0-20° C., comprising platelets; a compound according to any one of claims 1-9 or a pharma- ceutically acceptable salt thereof, or a combination thereof; and a physiologically acceptable carrier, wherein the compound is present at a concentration of 1 μM to 20 μM.

27. 27. The composition of claim 26, wherein the carrier is a buffer.

28. 27. The composition of claim 26, wherein the carrier is selected from platelet additive solution (PAS), saline, phosphate buffered saline, Tris buffered saline, Hank's buffered saline, water, or a combination thereof.

29. The composition of any one of claims 26 to 28, wherein the carrier comprises an electrolyte solution.

30. The composition of any one of claims 26 to 29, further comprising an additive.

31. NaCl, KCl, CaCl ２ , MgCl ２ , MgSO ４ , Sodium Citrate ３ , Citric Acid, NaHCO ３ 31. The composition of claim 30, wherein the additive is selected from sodium phosphate, sodium acetate, sodium gluconate, glucose, maltose, mannitol, and combinations thereof.

32. The composition of claims 30-31, wherein the additive is present in an amount of 0.5 mmol / L to 150 mmol / L.

33. 33. The composition of any one of claims 30 to 32, further comprising one or more components selected from D-ribose, D-glucose, Hank's solution, Hepes solution, bovine serum albumin, Tick anticoagulant peptide, and sterile water, or combinations thereof.

34. The composition of any one of claims 30 to 33, further comprising an additive selected from pH adjusting and buffering agents, tonicity adjusting agents, stabilizers, wetting agents, and combinations thereof.

35. The composition according to any one of claims 30 to 34, wherein the composition has a pH of from 5 to 8.

36. The composition of any one of claims 30 to 35, wherein the composition is isotonic.

37. A method for storing platelets, comprising storing said platelets in the presence of a compound according to any one of claims 1 to 9.

38. A method for storing platelets comprising storing said platelets in a composition according to any one of claims 26 to 36.

39. 39. The method of claim 38, wherein the platelets are stored for 10 to 14 days.

40. 39. The method of claim 38, wherein the storage is performed at a temperature between 0°C and 10°C.

41. 39. The method of claim 38, wherein the storage is performed at a temperature between 1°C and 6°C.

42. 1. A method for improving platelet survival during transfusion, comprising: contacting platelets with a compound or a pharma- ceutically acceptable salt thereof according to any one of claims 1 to 9, or a composition according to any one of claims 26 to 36, or with platelets stored according to a method according to any one of claims 37 to 41; and injecting the contacted platelets into a subject (except for a human).

43. 43. The method of claim 42, wherein the compound is present at a concentration of 1 μM to 20 μM.

44. 43. The method of claim 42, wherein the platelets are stored in the presence of the compound at a temperature between 1°C and 25°C.

45. 43. The method of claim 42, wherein the platelets are stored in the presence of the compound for 1 to 14 days.

46. 43. The method of claim 42, further comprising contacting the platelets with a physiologically acceptable carrier.

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