Prevention or treatment of cardiovascular disease with highly permeable prodrugs of aspirin and other NSAIDs

Abstract

Provided are 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, other highly permeable prodrugs of aspirin and other NSAIDs, and pharma- ceutically acceptable salts thereof, for use in and methods of preventing or treating cardiovascular diseases and conditions. Disclosed are pharmaceutical compositions, treatment kits, and devices comprising 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, other highly permeable prodrugs of aspirin and other NSAIDs, and pharma- ceutically acceptable salts thereof, as well as dosage forms, dosage amounts, and methods of their use by topical administration.

Description

[Technical field]

[0001] The present disclosure relates to the field of medical applications, in particular to the use of highly permeable prodrugs of aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs), and pharma- ceutically acceptable salts thereof, for the prevention or treatment of cardiovascular diseases or conditions, especially by topical or transdermal administration. [Background technology]

[0002] Aspirin has been used in humans and animals for over 100 years. Aspirin and other known NSAIDs, which have analgesic, anti-inflammatory, antipyretic, anti-inflammatory or antiplatelet properties, are widely used to treat various diseases, such as pain-related conditions such as cardiovascular diseases, inflammatory diseases and rheumatic diseases, headaches, migraines, toothaches, back pain, muscle pain, post-operative pain, etc.

[0003] Aspirin and other NSAIDs are usually administered orally to reach the site of action of pathology or disease. Unfortunately, the use of aspirin and other NSAIDs is associated with an increased risk of serious gastrointestinal (GI) toxicity, including gastroduodenal bleeding, gastric ulcers, gastritis, and GI perforation (see, for example, Non-Patent Document 1, Non-Patent Document 2, Non-Patent Document 3, Non-Patent Document 4, and Non-Patent Document 5). Furthermore, oral administration has limited effectiveness in preventing or treating certain diseases. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Cohn SM, et al., J. Clin. Invest. 1997; 99(6):1367-1379 [Non-Patent Document 2] Tarnawski AS and Ahluwalia A., Curr. Med. Chem. 2012; 19(1):16-27 [Non-Patent Document 3] Fries JF, J. Rheumatol Suppl. 1991; 28:6-10 [Non-Patent Document 4] Garcia Rodriguez LA, et al., Arch. Intern Med. 1998; 158(1):33-9 [Non-Patent Document 5] Richardson C, Emery P., Drug Saf. 1996; 15(4):249-60 Summary of the Invention [Problem to be solved by the invention]

[0005] Thus, there remains a need to develop new alternative delivery routes or methods that can realize the full efficacy of aspirin and other NSAIDs while avoiding GI toxicity. [Means for solving the problem]

[0006] The above-mentioned needs are met in the present disclosure by providing highly permeable prodrugs (HPPs) of aspirin and / or other NSAIDs, highly permeable compositions (HPCs) thereof, uses of the HPPs or HPCs, kits, therapeutic systems, dosage forms, devices using or including the HPPs or HPCs, and methods of treating various cardiovascular diseases and conditions.

[0007] In one aspect, the disclosure provides compounds of formula (I) and stereoisomers and pharma- ceutically acceptable salts thereof as HPPs of aspirin or analogs thereof, comprising a functional group covalently attached to a transport unit via a linker. JPEG2025504447000002.jpg6080

[0008] During the ceremony, L1 is O, S, NH, O-CH(L2), O-(CH2) n, O-CH(L2)-OC(=O), O-CH(L2)-O, S-CH(L2)-O, and -OC(=O)-, where n is an integer selected from 1 to 6; L2 at each occurrence is independently selected from H, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, and substituted and unsubstituted alkylamino; T is a transport unit that contains a protonatable amine group, e.g., a substituted or unsubstituted primary amine group, a substituted or unsubstituted secondary amine group, a substituted or unsubstituted tertiary amine group, or a substituted or unsubstituted heterocyclyl that contains a protonatable nitrogen;

[0009] R X is selected from H, 2,4-difluorophenyl, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl; R y is selected from H, substituted and unsubstituted alkylcarbonyl, substituted and unsubstituted alkoxycarbonyl, substituted and unsubstituted benzoyl, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl;

[0010] The compounds of formula (I) are capable of forming pharma- ceutically acceptable salts with acids via a protonatable transport unit T, which salts are capable of penetrating one or more biological barriers.

[0011] In one aspect, the present disclosure provides a pharmaceutical composition comprising an HPP of aspirin or an analog thereof characterized by formula (I) and a pharma- ceutically acceptable carrier. Such a pharmaceutical composition constitutes a highly permeable composition (HPC).

[0012] In one aspect, the present disclosure provides the use of HPP or HPC to prevent or treat various cardiovascular diseases or conditions through convenient local administration, which, among many other advantages, can generally overcome GI toxicity and unexpectedly improve efficacy of prevention or treatment compared to traditional oral administration.

[0013] In one aspect, the disclosure provides the use of HPP in the manufacture of a medicament for the prevention or treatment of various cardiovascular diseases or conditions, such as stroke, angina, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, and peripheral arterial disease.

[0014] In one aspect, the disclosure provides a method of preventing or treating a disease or condition, the method comprising administering to a subject a therapeutically effective amount of an HPP or HPC as disclosed herein. In some embodiments, the disease or condition is selected from stroke, angina, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, peripheral artery disease, and other cardiovascular diseases.

[0015] In one aspect, the disclosure provides a kit that includes an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride.

[0016] In one aspect, the disclosure provides a therapeutic system comprising a composition comprising an HPP of aspirin or other NSAIDs, such as 2-(diethylamino)ethyl 2-acetoxybenzoate, and / or a related highly permeable prodrug of aspirin or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride.

[0017] In one aspect, the disclosure provides a dosage form comprising a certain concentration of an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, for the treatment of cardiovascular disease.

[0018] In one aspect, the disclosure provides a device that can administer a certain unit dose of 2-(diethylamino)ethyl 2-acetoxybenzoate or a HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, to a subject suffering from a cardiovascular disease or condition.

[0019] The HPP or HPC disclosed herein can cross one or more biological barriers and can be administered locally (e.g., topically or transdermally) to reach locations where pathology occurs but cannot be adequately reached by existing drugs, thereby avoiding drug-associated gastrointestinal disorders or upper GI tract ulcer complications such as bleeding episodes, perforation, or gastric outlet obstruction, and unexpectedly improving the efficacy of prevention or treatment compared to oral administration. These and other advantages of the present disclosure will be better understood in view of the following detailed description, figures, examples, and claims. [Brief description of the drawings]

[0020] [Figure 1] FIG. 1 shows the effect of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride on cerebral infarct volume 14 days after middle cerebral artery occlusion (MCAO) (mean±SEM, assessed by TTC staining). [Diagram 2]FIG. 1 shows the effect of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride on body weight change in rats after MCAO. [Diagram 3] FIG. 1 shows the effect of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride on neurological deficit scores in rats after MCAO by daily assessment compared to the vehicle group. [Figure 4] FIG. 1 shows the effect of 2-(diethylamino)ethyl hydroxybenzoate hydrochloride on cerebral infarct volume 14 days after MCAO (mean±SEM, assessed by TTC staining).

[0021] [Diagram 5] FIG. 1 shows the effect of 2-(diethylamino)ethyl hydroxybenzoate hydrochloride on body weight change in rats after MCAO. [Figure 6] FIG. 1 shows the effect of 2-(diethylamino)ethyl hydroxybenzoate hydrochloride on neurological deficit scores in rats after MCAO by daily assessment compared to the vehicle group. [Figure 7] FIG. 1 shows the effect of (pyrrolidin-2-yl)methyl acetoxybenzoate hydrochloride on cerebral infarct volume 14 days after MCAO (mean±SEM, assessed by TTC staining). [Figure 8] FIG. 1 shows the effect of (pyrrolidin-2-yl)methyl acetoxybenzoate hydrochloride on body weight change in rats after MCAO.

[0022] [Figure 9] FIG. 1 shows the effect of (pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride on neurological deficit scores in rats after MCAO by daily assessment compared to the vehicle group. [Figure 10]FIG. 1 shows the effect of (pyrrolidin-2-yl)methyl 2′,4′-difluoro-4-acetoxy-[1,1′-biphenyl]-3-carboxylate hydrochloride on cerebral infarct volume 14 days after MCAO (mean±SEM, assessed by TTC staining). [Figure 11] FIG. 1 shows the effect of (pyrrolidin-2-yl)methyl 2′,4′-difluoro-4-acetoxy-[1,1′-biphenyl]-3-carboxylate hydrochloride on body weight change in rats after MCAO. [Figure 12] FIG. 1 shows the effect of (pyrrolidin-2-yl)methyl 2′,4′-difluoro-4-acetoxy-[1,1′-biphenyl]-3-carboxylate hydrochloride on neurological deficit scores in rats after MCAO by daily assessment compared to the vehicle group.

[0023] [Figure 13] FIG. 1 shows the effect of (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride on cerebral infarct volume 14 days after MCAO (mean±SEM, assessed by TTC staining). [Figure 14] FIG. 1 shows the effect of (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride on body weight change in rats after MCAO. [Figure 15] FIG. 1 shows the effect of (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride on neurological deficit scores in rats after MCAO by daily assessment compared to the vehicle group.

[0024] [Figure 16] FIG. 1 shows the effect of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride on neurological deficit scores in monkeys compared to vehicle and aspirin groups. [Figure 17]FIG. 1 shows the effect of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride on infarct volume in monkeys compared to vehicle and aspirin groups. [Figure 18] FIG. 1 shows the effect of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride compared to vehicle and aspirin groups on infarct volume in minipigs. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] In one embodiment, the present disclosure provides a method for the preparation of a transporter comprising the steps of: The present invention provides HPPs of aspirin or analogs thereof, characterized by JPEG2025504447000003.jpg6080 and stereoisomers and pharma- ceutically acceptable salts thereof.

[0026] During the ceremony, R X is selected from H, 2,4-difluorophenyl, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl; R y is selected from H, substituted and unsubstituted alkylcarbonyl, substituted and unsubstituted alkoxycarbonyl, substituted and unsubstituted benzoyl, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl, preferably R y is 2-acetoxybenzoyl or 2-hydroxybenzoyl.

[0027] L1 is O, S, NH, O-CH(L2), O-(CH2) n, O-CH(L2)-OC(=O), O-CH(L2)-O, S-CH(L2)-O, and -OC(=O)-, where n is an integer selected from 1 to 6; L2 at each occurrence is independently selected from H, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, and substituted and unsubstituted alkylamino.

[0028] T is a transport unit containing a protonatable amine group, e.g., a substituted or unsubstituted primary amine group, a substituted or unsubstituted secondary amine group, a substituted or unsubstituted tertiary amine group, or a substituted or unsubstituted heterocyclyl containing a protonatable nitrogen, and T is represented by Structure W-1, Structure W-2, Structure W-3, Structure W-4, Structure W-5, and Structure W-6: You can choose from JPEG2025504447000004.jpg91160.

[0029] R at each occurrence is independently selected from a bond, substituted and unsubstituted alkylene, substituted and unsubstituted cycloalkylene, substituted and unsubstituted heterocyclylene, substituted and unsubstituted alkenylene, substituted and unsubstituted alkynylene, substituted and unsubstituted arylene, and substituted and unsubstituted heteroarylene, where any CH2 in R can be optionally further replaced with O, S, or NR3, where R3 is hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, or C6-C7 alkyl. 10 Preferably, R is, at each occurrence, -CH2- or -CH2-CH2-.

[0030] R1 and R2 are independently selected from H, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted alkyloxyl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl, or alternatively, R1 and R2 together with the nitrogen atom to which they are attached form an optionally substituted heterocyclyl, which optionally further contains 1 or 2 additional heteroatoms independently selected from O, S, and N.

[0031] R 11 , R 12 , and R 13 are each independently a bond, an optionally substituted C1-C4 alkylene, or an optionally substituted C2-C4 alkenylene, wherein the alkylene and the alkenylene optionally have one CH2 group replaced by O, S, or NR3, and preferably R 11 , R 12 , and R 13 are each independently -CH2- or -CH2-CH2-.

[0032] In the above, any one of R1 and adjacent R in Structure W-2, Structure W-3, or Structure W-5 is 11 can be taken together with the nitrogen atom to which they are attached to form an optionally substituted heterocyclic ring, which optionally further contains one or two additional heteroatoms independently selected from O, S, and N; Here, the R in Structure W-2, Structure W-4, Structure W-5, or Structure W-6 11 and the above R 12 Or the above R 11 and the above R 13 are optionally connected by an optionally substituted alkylene bridge.

[0033] wherein HA is free and selected from pharma- ceutically acceptable acids, such as hydrochloride, hydrobromide, hydroiodide, nitric acid, sulfuric acid, bisulfate, phosphoric acid, phosphorous acid, phosphonic acid, isonicotinic acid, acetic acid, lactic acid, salicylic acid, citric acid, tartaric acid, pantothenic acid, bitartaric acid, ascorbic acid, succinic acid, maleic acid, gentisic acid, fumaric acid, gluconic acid, glucaronic acid, saccharic acid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and pamoic acid. However, the structure must form a stable compound without violating the principles of covalent bonding.

[0034] In some embodiments, when HA is absent, the compound of formula (I) is a free base. In some embodiments, in the compound of formula (I), R X is hydrogen, halogen, hydroxyl, thiol, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, or NR a R b where R a and R b are the same or different and independently hydrogen or C1 to C6 alkyl.

[0035] In some embodiments, in the compound of formula (I), R y is selected from hydrogen (H), CH3CO-, CH3CH2CO-, 2-hydroxybenzoyl, and 2-acetoxybenzoyl, where the phenyl portion of 2-hydroxybenzoyl or 2-acetoxybenzoyl is selected from halogen, hydroxyl, thiol, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, phenyl, and NR a R b and optionally further substituted with 1 to 3 substituents independently selected from a and R bare the same or different and are independently hydrogen or C1-C6 alkyl, where phenyl is selected from halogen, hydroxyl, thiol, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, and NR a R b and optionally substituted with 1 to 5, sometimes preferably 1 to 3, substituents independently selected from:

[0036] In some embodiments, in the compound of Formula (I), L is O, S, NH, or O(CH). n (n is 1, 2, 3, or 4). In some embodiments, in the compound of Formula (I), R is a bond or C1-C6 alkylene. In some embodiments, in the compound of Formula (I), T is structure W-1, where R1 and R2 are each hydrogen or C1-C6 alkyl.

[0037] In some embodiments, in the compound of Formula (I), T is Structure W-2, Structure W-3, Structure W-4, Structure W-5, or Structure W-6, where R is a bond or C1-C4 alkylene, R1 is hydrogen or C1-C6 alkyl, and R 11 is C1-C4 alkylene, R 12 and R 13 is independently a bond, CH, or CHCH.

[0038] In some embodiments, in compounds of Formula (I), T is a heterocyclyl selected from pyrrolidinyl, piperidinyl, piperazinyl, and morpholinyl. In some embodiments, R 11 , R 12 , and R 13 are each independently CH2, CH2CH2, CH=CH, CH2CH2CH 2、 CH=CHCH2, CH2CH2CH2CH 2、 CH2CH=CH-CH 2、 CH2CH2CH2CH2CH 2、CH2CH2CH2CH2CH2CH2, each of which is optionally substituted.

[0039] The present disclosure encompasses all reasonable combinations of the embodiments disclosed herein with respect to the structure of formula (I). For example, in some embodiments, in a compound of formula (I): R X is hydrogen, halogen, hydroxyl, thiol, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, or NR a R b where R a and R b are the same or different and independently hydrogen or C1 to C6 alkyl.

[0040] R y is selected from hydrogen (H), CH3CO-, CH3CH2CO-, 2-hydroxybenzoyl, and 2-acetoxybenzoyl, where the phenyl portion of 2-hydroxybenzoyl or 2-acetoxybenzoyl is selected from halogen, hydroxyl, thiol, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, phenyl, and NR a R b and optionally further substituted with 1 to 3 substituents independently selected from a and R b are the same or different and are independently hydrogen or C1-C6 alkyl, where phenyl is selected from halogen, hydroxyl, thiol, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, and NR a R b and optionally further substituted with 1 to 3 substituents independently selected from L1 is O, S, NH, or O(CH2) n (n is 1 or 2), and T is structure W-1, where R is a bond or C1-C4 alkylene, and R1 and R2 are each hydrogen or C1-C6 alkyl; or T is Structure W-2, Structure W-3, Structure W-4, Structure W-5, or Structure W-6, where R is a bond or C1-C4 alkylene, R1 is hydrogen or C1-C6 alkyl, and R 11 is C1-C4 alkylene, R 12 and R 13 is independently a bond, CH, or CHCH.

[0041] In some embodiments, the present disclosure provides a method for producing a pharmaceutical composition comprising: JPEG2025504447000005.jpg81161JPEG2025504447000006.jpg91156JPEG2025504447000007.jpg107157 or a pharma- ceutically acceptable salt thereof.

[0042] In some embodiments, the pharma- ceutically acceptable salt is formed with an acid selected from hydrochloride, hydrobromide, hydroiodide, nitric acid, sulfuric acid, bisulfate, phosphoric acid, phosphorous acid, phosphonic acid, isonicotinic acid, acetic acid, lactic acid, salicylic acid, citric acid, tartaric acid, pantothenic acid, bitartaric acid, ascorbic acid, succinic acid, maleic acid, gentisic acid, fumaric acid, gluconic acid, glucaronic acid, saccharic acid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and pamoic acid. In a preferred embodiment, the acid is hydrochloric acid.

[0043] In some embodiments, the functional unit in the compound of formula (I) (the salicylic acid moiety of aspirin) may be replaced by a moiety of another NSAID, such as diflunisal, acetyldiflunisal, salsalate, acetylsalsalate, and the like, replacing the hydroxyl on the carboxyl group of the NSAID.

[0044] In one aspect, the present disclosure provides the use of HPP or HPC to prevent or treat various cardiovascular diseases or conditions through convenient local administration, which can generally overcome GI toxicity, among many other advantages, compared to traditional oral administration.

[0045] In some embodiments, the present disclosure provides a method for the prevention or treatment of stroke, angina, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, peripheral artery disease, and other cardiovascular diseases, comprising administering to the patient a therapeutically effective amount of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, (pyrrolidin-2-yl)methyl acetoxybenzoate hydrochloride, 2-(diethylamino)ethyl hydroxybenzoate hydrochloride, 2-(pyrrolidin-2-yl)methyl hydroxybenzoate hydrochloride, 2-(diethylamino)ethyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride, (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride, or ... In one embodiment, the present invention provides for the use of aspirin and other NSAIDs such as 2-(diethylamino)ethyl 2',4'-difluoro-4-hydroxyl-[1,1'-biphenyl]-3-carboxylate hydrochloride, 2-(diethylamino)ethyl 2',4'-difluoro-4-hydroxyl-[1,1'-biphenyl]-3-carboxylate hydrochloride, (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-hydroxyl-[1,1'-biphenyl]-3-carboxylate hydrochloride, 2-(diethylamino)ethyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride, (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride, 2-(diethylamino)ethyl 2-(2-hydroxybenzoyl)oxybenzoate hydrochloride, (pyrrolidin-2-yl)methyl 2-(2-hydroxybenzoyl)oxybenzoate hydrochloride, or other HPPs or HPCs of aspirin and other NSAIDs.

[0046] In one aspect, the disclosure provides for the use of HPP in the manufacture of a medicament for the prevention or treatment of various cardiovascular diseases or conditions, such as stroke, angina, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, peripheral artery disease, atherosclerosis, and other cardiovascular diseases.

[0047] In one embodiment, the present disclosure relates to the use of 2-(diethylamino)ethyl 2-acetoxybenzoate and / or related highly permeable prodrugs of aspirin and / or other NSAIDs or pharma- ceutically acceptable salts thereof in the manufacture of a medicament.

[0048] In some embodiments, the present disclosure provides a method for the prevention or treatment of stroke, angina, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, peripheral artery disease, and other cardiovascular diseases, comprising administering to the patient a compound of formula (I) or (II) of formula (I) above, in the manufacture of a medicament for the prevention or treatment of stroke, angina, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, peripheral artery disease, and other cardiovascular diseases, comprising administering to the patient a compound of formula (I) above, in the manufacture of a medicament for the prevention or treatment of stroke, angina, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, peripheral artery disease, and other cardiovascular diseases ... and other uses of HPP or HPC of aspirin and other NSAIDs.

[0049] In some embodiments, the disclosure provides for the use of 2-(diethylamino)ethyl 2-acetoxybenzoate, 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, or other HPPs of aspirin in the manufacture of a medicament for the prevention or treatment of stroke, angina, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, peripheral artery disease, and other cardiovascular diseases.

[0050] In one aspect, the disclosure provides a method for preventing or treating stroke, angina, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, peripheral artery disease, and other cardiovascular diseases by local administration of an HPP or HPC of aspirin and / or other NSAIDs.

[0051] In one aspect, the disclosure provides a kit that includes an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride.

[0052] In one aspect, the disclosure provides a therapeutic system comprising a composition comprising an HPP of aspirin or other NSAIDs, such as 2-(diethylamino)ethyl 2-acetoxybenzoate, and / or a related highly permeable prodrug of aspirin or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride.

[0053] In one aspect, the disclosure provides a dosage form comprising a certain concentration of an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, for the treatment of cardiovascular disease.

[0054] In one aspect, the disclosure provides a device that can administer a certain unit dose of 2-(diethylamino)ethyl 2-acetoxybenzoate or a HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, to a subject suffering from a cardiovascular disease or condition.

[0055] In some embodiments, the disclosure provides a method of preventing or treating a cardiovascular disease or condition in a subject, comprising topically administering an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, to a site on the subject, particularly to one or more sites on the subject, in an amount of about 1 mg to about 1000 mg per administration, particularly 3 mg to 200 mg per administration.

[0056] In one aspect, the disclosure provides an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, for use in the prevention or treatment of a cardiovascular disease or condition in a subject, the HPP being administered locally to a site on the subject, particularly to one or more sites on the subject, in an amount of about 1 mg to about 1000 mg per dose, particularly 3 mg to 200 mg per dose.

[0057] In some embodiments, the disclosure provides for the use of an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, in the manufacture of a medicament for local administration to prevent or treat a cardiovascular disease or condition, wherein the HPP is administered locally to a site on a subject, particularly to one or more sites on a subject, in an amount of about 1 mg to about 1000 mg per dose, particularly 3 mg to 200 mg per dose.

[0058] In some embodiments, the disclosure provides a kit for preventing or treating a subject suffering from a cardiovascular disease or condition, comprising an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, administered locally to a site on the subject, particularly to one or more sites on the subject, in an amount of about 1 mg to about 1000 mg per dose, particularly 3 mg to 200 mg per dose.

[0059] In some embodiments, the disclosure provides a therapeutic system for preventing or treating a subject suffering from a cardiovascular disease or condition comprising a composition comprising an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, where the HPP is present as a free base or as a pharma- ceutically acceptable salt, wherein in the system the HPP is administered locally to a site on the subject, particularly to one or more sites on the subject, in an amount of about 1 mg to about 1000 mg per dose, particularly 3 mg to 200 mg per dose.

[0060] In some embodiments, the disclosure provides a method of preventing or treating a subject, comprising administering to a site on the subject an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, in a dose that is more than 1 cm of skin per dose, particularly to one or more sites on the subject. 2 Approximately 5μg per 1cm of skin 2 Approximately 3 mg per cm of skin 2 5μg per 1cm of skin 2 The method includes topically administering the compound in an amount of 3 mg per dose once daily, twice daily, three times daily, or four times daily.

[0061] In some embodiments, the present disclosure provides a HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, for use in the prevention or treatment of a subject, the HPP being administered to a site on the subject, particularly to one or more sites on the subject, up to 1 cm of skin per administration. 2 Approximately 5μg per 1cm of skin 2 Approximately 3 mg per cm of skin 2 5μg per 1cm of skin 2

[0023] The present invention provides HPP, administered topically in an amount of 3 mg per dose once daily, twice daily, three times daily, or four times daily.

[0062] In some embodiments, the skin area to which the drug is applied is about 5 cm 2 ~15000cm 2 , especially 25cm 2 ~5000cm 2 , especially 100cm 2 ~2500cm 2 It is.

[0063] In some embodiments, the disclosure provides for the use of an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, in the manufacture of a medicament, wherein the HPP in the medicament is administered to a site on a subject, particularly to one or more sites on a subject, in an amount of about 1 cm of skin per administration. 2 Approximately 5μg per 1cm of skin 2 Approximately 3 mg per cm of skin 2 5μg per 1cm of skin 2 The use is provided by topical administration in an amount of 3 mg per dose.

[0064] In some embodiments, the present disclosure provides a kit for preventing or treating a subject, comprising administering to a site on the subject, and in particular to one or more sites on the subject, up to 1 cm of skin per administration. 2 Approximately 5μg per 1cm of skin 2 Approximately 3 mg per cm of skin 2 5μg per 1cm of skin 2The kit includes a HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, administered topically in an amount of 3 mg per dose.

[0065] In some embodiments, the present disclosure provides a therapeutic system for preventing or treating a subject comprising a composition having an active ingredient HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, where HPP is present as a free base or as a pharma- ceutical acceptable salt, wherein the system provides the HPP to the subject, particularly to one or more sites on the subject, in a dose of less than 1 cm of skin per dose. 2 Approximately 5μg per 1cm of skin 2 Approximately 3 mg per cm of skin 2 5μg per 1cm of skin 2 The present invention provides a therapeutic system for topical administration in an amount of 3 mg per 100 mg of topical administration.

[0066] In some embodiments, the disclosure provides a dosage form, wherein the concentration of the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in the dosage form is from about 3 mg / mL to about 200 mg / mL, particularly 10 mg / mL to 100 mg / mL, or from about 3 mg / g to about 200 mg / g, particularly 10 mg / g to 100 mg / g.

[0067] In some embodiments, the present disclosure provides devices that can administer to a subject a unit dose of about 0.1 mg to about 30 mg, particularly 0.1 mg to 30 mg, of an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride.

[0068] In some embodiments, the present disclosure provides a propellant capable of nebulizing a unit dose of about 0.1 mg to about 30 mg, particularly 0.1 mg to 30 mg, of an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride.

[0069] In one aspect, the present disclosure is directed to assess the efficacy and safety of HPP of aspirin or other NSAIDs, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, when administered to the skin as a topical spray in subjects with cardiovascular disease.

[0070] In some embodiments, the present disclosure provides pharmaceutical compositions capable of penetrating biological barriers and methods of using the same to prevent or treat cardiovascular disease in humans and animals, particularly stroke, myocardial infarction, heart failure, angina pectoris, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, and peripheral arterial disease.

[0071] 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 belongs.

[0072] The term "cardiovascular disease" refers to a group of disorders of the heart and blood vessels, including: Coronary heart disease, i.e., disease of the blood vessels that supply the heart muscle; Cerebrovascular diseases, i.e. diseases of the blood vessels that supply the brain, Peripheral arterial disease, i.e., disease of the blood vessels that supply the arms and legs; Rheumatic heart disease, damage to the heart muscle and heart valves due to rheumatic fever caused by streptococcal bacteria; Congenital heart disease, i.e. birth defects that affect the normal development and functioning of the heart caused by malformations of the cardiac structure at birth, and Deep vein thrombosis and pulmonary embolism, i.e., a blood clot forms in the veins of the legs, which can break off and travel to the heart and lungs.

[0073] The term "alkyl" refers to a branched or unbranched monovalent aliphatic hydrocarbon group derived from an alkane by removing one hydrogen atom. In certain embodiments, the alkyl group contains 1 to 8 carbons. In certain embodiments, and sometimes preferably, the alkyl group contains 1 to 6 carbons, and in certain embodiments, and sometimes more preferably, the alkyl group contains 1 to 4 carbons. Examples of alkyl include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, and dodecyl. In certain embodiments, the alkyl group contains 1 to 12 carbons. The alkyl group can be substituted or unsubstituted.

[0074] The term "alkenyl" refers to any monovalent aliphatic hydrocarbon group derived by removing one hydrogen atom from an alkene. In certain embodiments, an alkenyl group contains 2 to 12 carbons. In certain embodiments, an alkenyl group contains 2 to 8 carbons. In certain embodiments, and sometimes preferably, an alkenyl group contains 2 to 6 carbons, and in certain embodiments, and sometimes more preferably, an alkenyl group contains 2 to 4 carbons. Examples of alkenyl include, but are not limited to, ethenyl, propenyl, butenyl, isobutenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, and the like. An alkenyl group can be substituted or unsubstituted.

[0075] The term "alkynyl" refers to a monovalent aliphatic hydrocarbon group derived by removing one hydrogen atom from an alkyne. In certain embodiments, an alkynyl group contains 2-12 carbons. In certain embodiments, an alkynyl group contains 2-8 carbons. In certain embodiments, and sometimes preferably, an alkynyl group contains 2-6 carbons, and in certain embodiments, and sometimes more preferably, an alkynyl group contains 2-4 carbons. Examples of alkynyl include, but are not limited to, ethynyl, propynyl, butynyl, isobutynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, undecynyl, dodecynyl, and the like. An alkynyl group can be substituted or unsubstituted.

[0076] The term "cycloalkyl" refers to any monovalent group formed by removing one hydrogen atom from a cycloalkane. In certain embodiments, a cycloalkyl group contains 3 to 10 carbons. In certain embodiments, a cycloalkyl group contains 3 to 8 carbons. In certain embodiments, and sometimes preferred, a cycloalkyl group contains 3 to 6 carbons. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. Cycloalkyls can be optionally substituted or unsubstituted.

[0077] The term "heterocyclyl" refers to a cycloalkyl in which at least one ring atom is a non-carbon atom. Examples of non-carbon ring atoms include, but are not limited to, S, O, and N. Representative examples of monocyclic heterocyclyls include, but are not limited to, pyrrolidyl, piperidyl, piperazinyl, morpholinyl, sulfomorpholinyl, homopiperazinyl, and the like.

[0078] The term "alkylene" refers to a saturated, straight or branched, divalent aliphatic hydrocarbon group derived from a parent alkane by removing two hydrogen atoms. The straight or branched chain groups contain 1 to 12 carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 carbon atoms), preferably 1 to 8 carbon atoms, more preferably 1 to 6 carbon atoms, and sometimes more preferably 1 to 4 carbon atoms. Non-limiting examples of alkylene groups include, but are not limited to, methylene (-CH-), 1,1-ethylene (-CH(CH)-), 1,2-ethylene (-CHCH-), 1,1-propylene (-CH(CHCH)-), 1,2-propylene (-CHCH(CH)-), 1,3-propylene (-CHCHCH-), 1,4-butylidene (-CHCHCHCH-), etc. Alkylene groups can be substituted or unsubstituted.

[0079] The term "alkenylene" refers to an alkylene as defined above having at least two carbon atoms and at least one carbon-carbon double bond, preferably C 2~12 Alkenylene, more preferably C 2~8 Alkenylene, sometimes more preferably C 2~6 Alkenylene, sometimes even more preferably C 2~4 Alkenylene. Non-limiting examples of alkenylene groups include, but are not limited to, -CH=CH-, -CH=CHCH2-, -CH=CHCH2CH2-, -CH2CH=CHCH2-, etc. Alkenylene groups can be substituted or unsubstituted.

[0080] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or polycyclic fused ring (a "fused" ring system means that each ring in the system shares an adjacent pair of carbon atoms with another ring in the system) group having a completely conjugated pi-electron system. Preferably, the aryl is 6- to 10-membered, such as phenyl and naphthyl, most preferably phenyl. Aryl groups can be substituted or unsubstituted.

[0081] The term "heteroaryl" refers to a 5-14 membered aryl system having 1-4 heteroatoms selected from O, S, and N as ring atoms. Preferably, the heteroaryl is 5-10 membered (5-, 6-, 7-, 8-, 9-, and 10-membered, etc.), more preferably 5- or 6-membered, such as thiadiazolyl, pyrazolyl, oxazolyl, oxadiazolyl, imidazolyl, triazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. Heteroaryls may be fused with aryl, heterocyclyl, or cycloalkyl rings, where the ring attached to the parent structure is a heteroaryl. Heteroaryl groups may be substituted or unsubstituted.

[0082] The term "alkoxy" refers to any -O-(alkyl), such as methoxy, ethoxy, propoxy, butoxy, and the like.

[0083] The term "cycloalkoxy" refers to -O-(cycloalkyl), for example, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.

[0084] The term "bond" refers to a covalent bond using the "-" symbol. The term "hydroxyl" refers to an --OH group. The term "halogen" refers to a fluorine, chlorine, bromine, or iodine atom. The term "amino" refers to the group --NH. The term "alkylthio" refers to alkyl-S-.

[0085] The term "alkylamino" refers to "alkyl-NH-" or sometimes dialkylamino (-NR a R b ), where R a and R b) may be the same or different. Sometimes preferably, the alkyl group is a C1-C6 alkyl, and sometimes more preferably, the alkyl is a C1-C4 alkyl. Examples of alkylamino include, but are not limited to, CH3-NH-, -N(CH3)2, -N(CH2CH3)2, -NHCH2CH3, -N(CH3)(CH2CH3), -NH-Bu. t , -N(CH3)(Bu t ) etc.

[0086] The term "cyano" refers to the group -CN. The term "haloalkyl" means an alkyl group substituted with one or more halogen atoms, where the halogen atoms can be the same or different. The term "nitro" refers to the group --NO.sub.2. The term "oxo" refers to the group =O. The term "carboxyl" refers to the group -C(O)OH. The term "alkoxycarbonyl" refers to the group -C(O)O(alkyl). The term "alkylcarbonyl" refers to a -C(O)-alkyl group.

[0087] The term "optional" or "optionally" means that the subsequently described event or circumstance may, but does not necessarily, occur, and the description includes cases where the event or circumstance may or may not occur. For example, "a heterocyclyl group optionally substituted with alkyl" means that an alkyl group may, but does not necessarily, be present, and the description includes cases where the heterocyclyl group is substituted with alkyl and cases where the heterocyclyl group is not substituted with alkyl.

[0088] The term "substituted" refers to one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, being independently replaced with a corresponding number of substituents. A person skilled in the art can determine whether or not substitution is possible without undue effort through experiment or theory. For example, the combination of an amino group or hydroxyl group having free hydrogen and a carbon atom having an unsaturated bond (such as an olefin) may be unstable.

[0089] The term "covalent bond principle" as used herein refers to the basic rules and principles in the formation of covalent bonds in organic compounds as generally understood by those skilled in the art. For example, carbon atom is tetravalent and can only form four covalent bonds (e.g., four single bonds, or one double bond plus two single bonds, etc.), and oxygen is divalent and can only form two covalent bonds (two single bonds at -O-, or one double bond at =O).

[0090] The term "prodrug" refers to a compound that can be transformed in vivo, such as by hydrolysis in blood, to yield the parent compound that is active under physiological conditions. Common examples include, but are not limited to, ester and amide forms of a compound having an active form bearing a carboxylic acid moiety. In particular, the present disclosure provides a unique class of prodrugs, so-called "highly permeable prodrugs", as defined in the present disclosure.

[0091] When any group in any HPP structure is indicated as being either "substituted" and / or "unsubstituted," this means that the group may be optionally substituted with one or more, preferably one to five, and sometimes more preferably one to three, substituents independently selected from halogen, cyano, nitro, amino, alkyl, haloalkyl, alkoxy, haloalkoxy, aryl, alkylthio, alkylamino, alkylsulfonyl (alkylsulfone), alkylsulfoxyl (alkylsulfoxide), acyloxy, carboxylic acid, carboxylic acid ester, and carboxamide groups, and the like. Alkyl groups can have 1 to 10 carbon atoms, sometimes preferably 1 to 6 carbon atoms, and sometimes more preferably 1 to 4 carbon atoms. Esters can have C1 to C6 carbon atoms, ... 10 It may be an ester of an alcohol, sometimes preferably a C1 to C6 alcohol, sometimes more preferably a C1 to C4 alcohol.

[0092] In some embodiments, when an alkyl group, an alkenyl group, an alkynyl group, a cycloalkyl group, a heterocyclyl group, an aryl group, a heteroaryl group, or the like, or a portion thereof is substituted, the substituent(s) may be substituted at any available point of attachment(s), and the substituents may be selected from the group consisting of a C1-C6 alkyl group, a halogen group, a C1-C6 alkoxy group, a C1-C6 alkenyl group, a C1-C6 alkynyl group, a C1-C6 alkylthio group, a C1-C6 alkylamino group, a di-(C1-C6 alkyl)amino group, a thiol group, a hydroxyl group, a nitro group, a cyano group, an amino group, a C3-C6 cycloalkyl group, a 5-membered to 10-membered heterocyclyl group, a C6-C 10There may be one or more, sometimes preferably one to five, sometimes more preferably one to three groups independently selected from an aryl group, a 5-membered to 10-membered heteroaryl group, a C3-C6 cycloalkoxy group, a C1-C6 cycloalkylthio group, a 5-membered to 10-membered heterocyclylthio group, and an oxo group. In some embodiments, sometimes preferably, the substituents are independently selected from a C1-C6 alkyl group, a halogen group, a C1-C6 alkoxy group, a C1-C6 alkylthio group, a C1-C6 alkylamino group, a di-(C1-C6 alkyl)amino group, a thiol group, a hydroxyl group, a nitro group, a cyano group, an amino group, and an oxo group. In some embodiments, sometimes more preferably, the substituents are independently selected from a C1-C4 alkyl group, a halogen group, a C1-C4 alkoxy group, a C1-C4 alkylthio group, a C1-C4 alkylamino group, a di-(C1-C4 alkyl)amino group, a thiol group, a hydroxyl group, a nitro group, a cyano group, an amino group, and an oxo group. As would be understood by one of ordinary skill in the art, an oxo (=O) group cannot be a substituent on an aryl or heteroaryl group or on an unsaturated carbon in any other group.

[0093] As used in the specification and claim(s), the words "comprising" (and all forms of "comprising" such as "comprise" and "comprises"), "having" (and all forms of "having" such as "have" and "has"), "including" (and all forms of "including" such as "includes" and "include"), or "containing" (and all forms of "containing" such as "contains" and "contain") are inclusive, i.e., open-ended, and do not exclude further, unrecited elements or method steps.

[0094] The terms "a" and "an" and "the" and similar designations in the context of describing the present invention (particularly in the context of the appended claims) should be construed to cover both the singular and the plural, unless otherwise specified herein or clearly contradicted by context. The use of the word "a" or "an" when used in conjunction with the word "comprising" in the claims and / or specification may mean "one," but is also consistent with the meaning of "one or more," "at least one," and "one or more than one."

[0095] Where the plural is used for compounds, salts and the like, this is taken to mean also a single compound, salt or the like.

[0096] As used herein, the term "and / or" refers to and includes any and all possible combinations of one or more of the associated listed items. When used in a list of two or more items, the term "and / or" means that any one of the listed items can be used alone, or any combination of two or more of the listed items can be used. For example, if a composition, combination, configuration, juxtaposition, or group is described as including (or comprising) component A, component B, component C, and / or component D, the composition can include A alone, B alone, C alone, D alone, a combination of A and B, a combination of A and C, a combination of A and D, a combination of B and C, a combination of B and D, a combination of C and D, a combination of A and B and C, a combination of A and B and D, a combination of A and C and D, a combination of B and C and D, or a combination of A and B and C and D.

[0097] Throughout this application, the term "about" or "approximately" is used to indicate that a value includes the inherent variation of error for the device, method used to determine the value, or the variation that exists between study subjects. In one embodiment, the term "about" or "approximately" generally means within 10%, particularly within 9%, particularly within 8%, particularly within 7%, particularly within 6%, particularly within 5%, particularly within 4%, particularly within 3%, particularly within 2%, particularly within 1%, and particularly within 0.5% of a given value or range.

[0098] The terms "treat", "treating" or "treatment" as used herein include a treatment or therapeutic regimen that alleviates, reduces or relieves at least one symptom in a subject or results in a delay in the progression of a proliferative disorder. For example, a treatment can be a decrease in one or several symptoms of a disorder, or a complete eradication of a disorder, such as stroke, myocardial infarction, and / or cardiovascular disease. Within the meaning of this disclosure, the term "treat" also means arresting, delaying the onset (i.e., the period prior to clinical manifestation of a disorder), and / or reducing the risk of developing or worsening a disorder.

[0099] In some embodiments, the term "dose" as used herein refers to the amount of drug or active ingredient taken by an individual subject at any one time, particularly the total amount of drug or active ingredient taken by an individual subject at any one time per site.

[0100] In some embodiments, the term "dosage form" as used herein refers to a unitary administration of an active agent. Examples of dosage forms include tablets, capsules, injections, suspensions, solutions, emulsions, creams, ointments, suppositories, inhalable forms, transdermal forms, etc.

[0101] In some embodiments, the term "unit dose" or "dosage unit" refers to a dosage form that is configured to deliver a specified amount or dose of a composition or its components. Examples of dosage forms for topical administration include, but are not limited to, transdermal patches, creams, foams, gels, lotions, ointments, pastes, powders, shake lotions, solids, sponges, tapes, tinctures, vapors, injections, drops, rinses, sprays, and liquids. A "unit dose" or "dosage unit" may be configured to provide a complete unit dose or a portion thereof (e.g., 1 / 2, 1 / 3, or 1 / 4 of a dose). The predetermined amount in each unit dose may depend on factors including, but not limited to, the unique characteristics of the active compound and the particular therapeutic effect to be achieved, as well as the limitations inherent in the technology of making and administering such unit doses. For example, the unit dose may be a transdermal patch, a spray, i.e., a single spray in a spray application, a drop in a drop application, a certain length of tape, a rice grain or pea-sized ointment, or a scoop or spoonful of ointment. A unit dose measuring device, such as a cup, scoop, syringe, dropper, spoon, or irrigation device, may hold a measured amount of the composition equivalent to a dosage form, such as a cream, foam, gel, lotion, ointment, paste, powder, shake lotion, and solid, a full unit dose or a portion thereof (e.g., 1 / 2, 1 / 3, or 1 / 4 of a dose). There may be a single unit dose or multiple unit doses in a single administration dosage. The kit may include instructions regarding the size of the unit dose or a portion thereof.

[0102] The term "pharmacologically acceptable" is defined herein to refer to those compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for contact with the tissues of a subject without undue toxicity, irritation, allergic response, and other problematic complications, commensurate with a reasonable benefit / risk ratio.

[0103] The term "pharmaceutical composition" is defined herein to refer to a substance or mixture or solution containing at least one therapeutic agent that is administered to a subject to prevent or treat, particularly to treat, a particular disease or condition affecting the subject.

[0104] The term "other NSAIDs" as used herein refers to any NSAID other than aspirin, particularly salicylic acid, diflunisal, acetyldiflunisal, salsalate, and acetylsalsalate.

[0105] The term "pharmaceutical acceptable salt" refers to a salt of a compound of the present invention that is safe to administer to a subject. For a review of pharmaceutical acceptable salts, see Berge, et al., J. Pharm. Sci., 1977, 66, 1-19, which is incorporated herein by reference.

[0106] It is understood that the therapeutic agent may be administered daily in a single unit dose or in multiple unit doses, and / or may be administered daily in a single dose (once per day, qd) or in divided doses (two or more times per day, e.g., twice per day, bid).

[0107] The term "day" as used herein refers to either one calendar day or one 24-hour period in any time zone.

[0108] The term "patient" or "subject" is intended to include animals, including warm-blooded animals. Examples of patients include mammals, such as humans, dogs, cows, horses, pigs, sheep, goats, cats, mice, rabbits, rats, and transgenic non-human animals. In some embodiments, the patient is a human, e.g., a human suffering from, at risk of, or potentially susceptible to a disease, e.g., suffering from stroke, myocardial infarction, and / or cardiovascular disease.

[0109] In some embodiments, the term "transdermal administration" refers to administration of a transdermal dose, unit dose, or dosage form. The term "transdermally administering" refers to administering a transdermal dose, unit dose, or dosage form. The term "transdermally administered" refers to administering by a transdermal dose, unit dose, or dosage form. A patient and / or subject being "transdermally administered" is equivalent to the patient and / or subject receiving a "transdermal administration." "Administering transdermally" to a patient and / or subject is equivalent to administering a "transdermal administration" to the patient and / or subject.

[0110] In some embodiments, the term "site" (of a subject) refers to the human organ or body where a disease is found, for example the skin and / or body surface near the heart, brain, lungs, head, neck, chest, arms, legs, and / or back, which are themselves suffering from a disease, in particular poor blood flow, blood clotting, tissue inflammation, and / or cell death, in particular blood clotting, vascular inflammation, and / or tissue inflammation, such as the brain, heart, lungs, other organs and / or other tissues, more particularly stroke, myocardial infarction, and / or other cardiovascular disease.

[0111] In some embodiments, correspondingly, the term "administering to a site" (of a subject) means administering to (a) a location on the skin and / or body surface in the vicinity corresponding to or proximate to that "site," and / or (b) a location on the skin and / or body surface in the vicinity that provides a route by which that "site" can be reached.

[0112] For example, the site may be the skin proximate to a location that is suffering from, at risk of suffering from, or likely to suffer from a disease, such as the brain, heart, lungs, and / or other organs, more particularly a condition such as stroke, myocardial infarction, and / or other cardiovascular disease. Administration to the site may be administration to the skin and / or proximate to a body surface. Administration to the site may be within about 1 cm to about 20 cm, particularly within about 5 cm to about 15 cm, particularly about 1 cm, about 2 cm, about 3 cm, about 4 cm, about 5 cm, about 6 cm, about 7 cm, about 8 cm, about 9 cm, about 10 cm, about 11 cm, about 12 cm, about 13 cm, about 14 cm, about 15 cm, and / or from the environment at a distance of about 1 cm to about 100 cm, particularly about 5 cm to about 50 cm, particularly about 5 cm, about 10 cm, about 15 cm, about 20 cm, about 25 cm, about 30 cm, about 35 cm, about 40 cm, about 45 cm, about 50 cm, about 40 cm, about 45 cm, about 50 cm in any direction around the brain, heart, lungs, and / or other organs.

[0113] In some embodiments, the term "close" or "near" refers to within about 1 cm to about 100 cm, particularly within about 10 cm to about 50 cm, and particularly within a distance selected from about 10 cm, about 15 cm, about 20 cm, about 30 cm, about 35 cm, about 40 cm, about 45 cm, and about 50 cm, of the center of the site, i.e., the brain, heart, lungs, and / or other organs.

[0114] In some embodiments, the term "symptom" refers to any condition such as disease, inflammation, chest pain, palpitations, malaise, fever, shortness of breath, extreme fatigue, angina, leg and / or arm pain, swelling, fatigue, fainting, headache, arm and / or leg weakness, facial muscle weakness, speech impairment, loss of vision, coordination problems, dizziness, loss of consciousness, etc. In particular, symptoms can include chest pain, shortness of breath, headache, dizziness, arm and / or leg weakness, loss of consciousness associated with blood clots, particularly stroke, myocardial infarction, and / or other cardiovascular diseases.

[0115] In some of the above-described embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered in an amount of about 1 mg to about 1000 mg, particularly about 3 mg to about 210 mg, and especially about 35 mg to about 140 mg per dose, once daily, twice daily, three times daily, or four times daily, and sometimes preferably twice daily.

[0116] For example, 3.5mg, 7mg, 10.5mg, 14mg, 17.5mg, 21mg, 24.5mg, 28mg, 31.5mg, 35mg, 38.5mg, 42mg, 45.5mg, 49mg, 52.5mg, 56mg, 59.5mg, 63mg, 66.5mg, 70mg, 73.5mg, 77mg, 80.5mg, 84mg, 87.5mg, 91mg, 100mg, 120mg, 140mg, 160mg, 180mg, 190mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg, 340mg, 350mg, 360mg, 370mg, 380mg, 390mg, 400mg, 410mg, 420mg, 45.5mg, 49mg, 52.5mg, 56mg, 59.5mg, 63mg, 66.5mg, 70mg, 73.5mg, 77mg, 80.5mg, 84mg, 87.5mg, 91mg, 100mg, 110mg, 120mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 180mg, 190mg, 190mg, 180mg, 190mg, 190mg, 190mg, mg, 94.5 mg, 98 mg, 101.5 mg, 105 mg, 108.5 mg, 112 mg, 115.5 mg, 119 mg, 122.5 mg, 126 mg, 129.5 mg, 133 mg, 136.5 mg, 140 mg, 143.5 mg, 147 mg, 150.5 mg, 154 mg, 157.5 mg, 161 mg, 164.5 mg, 168 mg, 171.5 mg, 175 mg, 178.5 mg, 182 mg, 185.5 mg, 189 mg, 192.5 mg, 196 mg, 199.5 mg, 203 mg, 206.5 mg, and 210 mg can be administered in once daily or twice daily amounts.

[0117] In some of the above embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered to 1 cm of skin per dose. 2 Approximately 5μg per 1cm of skin 2 Approximately 15 mg per cm of skin 2 Approximately 10μg per 1cm of skin 2 Approximately 7 mg per cm of skin 2 Approximately 30μg per 1cm of skin 2 Approximately 2 mg per cm of skin 2 Approximately 50μg per 1cm of skin2 Approximately 1.5 mg per cm of skin 2 Approximately 70μg per 1cm of skin 2 Approximately 1 mg per cm of skin, sometimes preferably 2 Approximately 100μg per 1cm of skin 2 approximately 700 μg per cm of skin, and sometimes more preferably 2 Approximately 150μg per 1cm of skin 2 It is administered in an amount of approximately 500 μg per dose.

[0118] For example, 17.5 μg / cm per dose 2 , 35μg / cm 2 , 70μg / cm 2 , 140μg / cm 2 , 280μg / cm 2 , 560μg / cm 2 , 700μg / cm 2 , 1 mg / cm 2 , 2 mg / cm 2 , 3.5 mg / cm 2 , or 7 mg / cm 2 It can be administered in cutaneous amounts.

[0119] In some of the above embodiments, the subject is a warm-blooded animal. In some of the above embodiments, the subject is a mammal. In some of the above embodiments, the subject is a primate. In some of the above embodiments, the subject is a human. In some of the above embodiments, the subject is a minor. In some of the above embodiments, the subject is a minor, and the minor is under the age of 16. In some of the above embodiments, the subject is a human adult. In some of the above embodiments, the adult is 16 years of age or older.

[0120] In some of the above embodiments, the subject is a subject suffering from, at risk of, or potentially susceptible to suffering from the condition, and / or the medicament is for use in a subject suffering from, at risk of, or potentially susceptible to suffering from the condition. In some of the above embodiments, the subject is a subject suffering from, at risk of, or potentially susceptible to a condition, in particular a subject suffering from, at risk of, or potentially susceptible to a cardiovascular disease or condition such as stroke, angina pectoris, myocardial infarction, heart failure, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, peripheral arterial disease, etc., and / or the medicament is for a subject suffering from, at risk of, or potentially susceptible to a condition, in particular a subject suffering from, at risk of, or potentially susceptible to a cardiovascular disease or condition such as stroke, angina pectoris, myocardial infarction, heart failure, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, peripheral arterial disease, etc.

[0121] In some of the above embodiments, the subject is a subject suffering from, at risk of, or potentially susceptible to suffering from, a blood clot (clotting), and / or the medicament is for use in a subject suffering from, at risk of, or potentially susceptible to suffering from, a blood clot (clotting).

[0122] In some of the above embodiments, the subject is a subject suffering from, at risk of suffering from, or potentially capable of suffering from, a disease, inflammation, chest pain, palpitations, numbness, fever, shortness of breath, excessive fatigue, angina, pain in the legs and / or arms, swelling, fatigue, fainting, headache, weakness in the arms and / or legs, weakness of the facial muscles, difficulty in speaking, loss of vision, coordination problems, dizziness, loss of consciousness, and / or the medicament is for a subject suffering from, at risk of suffering from, or potentially capable of suffering from, a disease, inflammation, chest pain, palpitations, numbness, fever, shortness of breath, excessive fatigue, angina, pain in the legs and / or arms, swelling, fatigue, fainting, headache, weakness in the arms and / or legs, weakness of the facial muscles, difficulty in speaking, loss of vision, coordination problems, dizziness, loss of consciousness.

[0123] In some of the above-described embodiments, the site of the subject includes one or more surfaces. In some of the above-described embodiments, the site of the subject includes its neck surface, chest surface, back surface, lower back surface, head surface, cheek surface, shoulder surface, arm surface, hand surface, leg surface, and / or abdominal surface.

[0124] In some of the above embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered topically to one or more of the neck, chest, back, lower back, head, cheek, shoulder, arm, hand, leg, and / or abdominal surfaces.

[0125] In some of the above-mentioned embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered locally by transdermal administration. In some of the above-mentioned embodiments, the HPP is administered locally by a dosage form selected from one or more of transdermal patches, creams, foams, gels, lotions, ointments, pastes, powders, shake lotions, solids, sponges, tapes, tinctures, vapors, injections, drops, rinses, sprays, and liquids. In some of the above-mentioned embodiments, the HPP is administered locally by a dosage form selected from one or more of transdermal drops, rinses, and sprays. In some of the above-mentioned embodiments, the HPP is administered locally by a spray. In some of the above-mentioned embodiments, the HPP is administered locally by a spray to a subject suffering from stroke, myocardial infarction, and / or cardiovascular disease. In some of the above-described embodiments, the HPP is administered locally by drops. In some of the above-described embodiments, the HPP is administered locally by drops to a subject suffering from stroke, myocardial infarction, and / or cardiovascular disease.

[0126] In some of the above embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered topically in a dosage form containing one or more unit doses. In some of the above embodiments, the dosage form may be selected from one or more of a transdermal patch, cream, foam, gel, lotion, ointment, paste, powder, shake lotion, solid, sponge, tape, tincture, vapor, injection, drop, rinse, spray, and liquid, and the dosage form may contain one or more unit doses.

[0127] In some of the above-mentioned embodiments, the dosage form is a spray application. In some of the above-mentioned embodiments, the dosage form is a spray to a subject suffering from stroke, myocardial infarction, and / or cardiovascular disease. In some of the above-mentioned embodiments, the dosage form is a multiple spray, and each unit dose is a single spray in the multiple sprays. In some of the above-mentioned embodiments, the dosage form is a multiple patch, and each unit dose is a single patch in the multiple patches. In some of the above-mentioned embodiments, the dosage form is a drop application. In some of the above-mentioned embodiments, the dosage form is a drop to a subject suffering from stroke, myocardial infarction, and / or cardiovascular disease. In some of the above-mentioned embodiments, the dosage form is a multiple drop, and each unit dose is a single drop in the multiple drops.

[0128] In some of the above-described embodiments, a composition comprising an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered topically to a subject. In some of the above-described embodiments, a unit dose comprising a composition comprising 2-(diethylamino)ethyl acetoxybenzoate hydrochloride is administered topically to a subject.

[0129] In some of the above embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered topically dissolved in a solution. In some of the above embodiments, the composition is a solution. In some of the above embodiments, the composition is an alcoholic solution. In some of the above embodiments, the composition is an acetone solution. In some of the above embodiments, the composition is a dimethylsulfoxide solution. In some of the above embodiments, the composition is an aqueous alcohol solution. In some of the above embodiments, the composition is an aqueous acetone solution. In some of the above embodiments, the composition is an aqueous dimethylsulfoxide solution. In some of the above-described embodiments, the composition is a solution comprising water and an alcohol, wherein the alcohol is at least one, at least two, or more selected from the group consisting of methanol, ethanol, propanol, isopropanol, n-butanol, isobutanol, tert-butanol, n-amyl alcohol, isoamyl alcohol, activated amyl alcohol, tert-amyl alcohol, neopentyl alcohol, methyl n-propyl carbinol, methyl isopropyl carbinol, and 3-pentanol. In some of the above-described embodiments, the composition is a solution comprising water and ethanol and / or isopropanol.

[0130] In some of the above-mentioned embodiments, the composition is an aqueous ethanol solution. In some of the above-mentioned embodiments, the composition is an aqueous ethanol solution of 0% (volume / volume) to 75% (volume / volume). In some of the above-mentioned embodiments, the composition is an aqueous ethanol solution of 10% (volume / volume), 15% (volume / volume), 20% (volume / volume), 25% (volume / volume), 30% (volume / volume), 35% (volume / volume), 40% (volume / volume), 45% (volume / volume), 50% (volume / volume), 55% (volume / volume), 60% (volume / volume), 65% (volume / volume), 70% (volume / volume), or 75% (volume / volume). In some of the above-mentioned embodiments, the composition is an aqueous ethanol solution of 15% (volume / volume).

[0131] In some of the above-mentioned embodiments, the concentration of HPP in the composition is about 5 mg / mL to about 500 mg / mL, particularly 5 mg / mL to 500 mg / mL, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, or other NSAID. In some of the above-mentioned embodiments, the concentration of HPP in the composition is about 30 mg / mL to about 150 mg / mL, particularly 30 mg / mL to 150 mg / mL. In some of the above-mentioned embodiments, the concentration of HPP in the composition is about 50 mg / mL to about 100 mg / mL, particularly 50 mg / mL to 100 mg / mL. In some of the above-mentioned embodiments, the concentration of HPP in the composition is about 60 mg / mL to about 90 mg / mL, particularly 60 mg / mL to 90 mg / mL. In some of the above-described embodiments, the concentration of HPP in the composition is about 75 mg / mL to about 85 mg / mL, particularly 75 mg / mL to 85 mg / mL. In some of the above-described embodiments, the concentration of HPP in the composition is about 79 mg / mL, particularly 79 mg / mL.

[0132] In some of the above-mentioned embodiments, the volume of the composition in the unit dose is about 0.01 mL to about 1 mL, particularly 0.01 mL to 1 mL. In some of the above-mentioned embodiments, the volume of the composition in the unit dose is about 0.03 mL to about 0.3 mL, particularly 0.03 mL to 0.3 mL. In some of the above-mentioned embodiments, the volume of the composition in the unit dose is about 0.05 mL to about 0.2 mL, particularly 0.05 mL to 0.2 mL. In some of the above-mentioned embodiments, the volume of the composition in the unit dose is about 0.05 mL to about 0.15 mL, particularly 0.05 mL to 0.15 mL. In some of the above-mentioned embodiments, the volume of the composition in the unit dose is about 0.1 mL, particularly 0.1 mL.

[0133] In some of the above-mentioned embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered in an amount of about 0.1 mg to about 20 mg, particularly 0.1 mg to 20 mg, per unit dose. In some of the above-mentioned embodiments, the HPP is administered in an amount of about 1 mg to about 18 mg, particularly 1 mg to 18 mg, per unit dose. In some of the above-mentioned embodiments, the HPP is administered in an amount of about 3 mg to about 16 mg, particularly 3 mg to 16 mg, per unit dose. In some of the above-mentioned embodiments, the HPP is administered in an amount of about 5 mg to about 15 mg, particularly 5 mg to 15 mg, per unit dose. In some of the above-mentioned embodiments, the HPP is administered in an amount of about 6 mg to about 12 mg, particularly 6 mg to 12 mg, per unit dose. In some of the above-described embodiments, HPP is administered in an amount of about 7 mg to about 10 mg, particularly 7 mg to 10 mg, per unit dose. In some of the above-described embodiments, HPP is administered in an amount of about 7.5 mg to about 9 mg, particularly 7.5 mg to 9 mg, per unit dose.

[0134] In some of the above-described embodiments, one or more unit doses are topically administered to a subject in a single dose, the one or more unit doses being a 1 unit dose, 2 unit doses, 3 unit doses, 4 unit doses, 5 unit doses, 6 unit doses, 7 unit doses, 8 unit doses, 9 unit doses, 10 unit doses, 11 unit doses, 12 unit doses, 13 unit doses, 14 unit doses, 15 unit doses, 16 unit doses, 17 unit doses, 18 unit doses, 19 unit doses, 20 unit doses, 21 unit doses, 22 unit doses, 23 unit doses, 24 unit doses, 25 unit doses, 26 unit doses, 27 unit doses, 28 unit doses, 29 unit doses, 30 unit doses, 31 unit doses, 32 unit doses, 33 unit doses, 34 unit doses, 35 unit doses, 36 unit doses, 37 unit doses, 38 unit doses, 39 unit doses, 40 unit doses, 41 unit doses, 42 unit doses, 43 unit doses, 44 unit doses, 45 unit doses, 46 unit doses, 47 unit doses, 48 ​​unit doses, 49 unit doses, 50 unit doses, 51 unit doses, 52 unit doses, 53 unit doses, 54 unit doses, 55 unit doses, 56 unit doses, 57 unit doses, 58 unit doses, 59 unit doses, 60 unit doses, 61 unit doses, 62 unit doses, 63 unit doses, 64 unit doses, 65 unit doses, 66 unit doses, 67 unit doses, 68 unit doses, 69 unit doses, 7 The dosage form is selected from the group consisting of 1 unit dose, 24 unit dose, 25 unit dose, 26 unit dose, 27 unit dose, 28 unit dose, 29 unit dose, 30 unit dose, 31 unit dose, 32 unit dose, 33 unit dose, 34 unit dose, 35 unit dose, 36 unit dose, 37 unit dose, 38 unit dose, 39 unit dose, 40 unit dose, 41 unit dose, 42 unit dose, 43 unit dose, 44 unit dose, 45 unit dose, 46 unit dose, 47 unit dose, 48 unit dose, 49 unit dose and 50 unit dose.

[0135] In some of the above embodiments, between 5 unit doses and 30 unit doses are administered locally to the subject in a single administration. In some of the above embodiments, between 10 unit doses and 20 unit doses are administered locally to the subject in a single administration. In some of the above embodiments, a 10 unit dose is administered locally to the subject in a single administration. In some of the above embodiments, a 15 unit dose is administered locally to the subject in a single administration. In some of the above embodiments, a 20 unit dose is administered locally to the subject in a single administration. In some of the above embodiments, a 25 unit dose is administered locally to the subject in a single administration. In some of the above embodiments, a 30 unit dose is administered locally to the subject in a single administration.

[0136] In some of the above embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered by a propellant capable of delivering about 0.1 mg to about 20 mg, particularly 0.1 mg to 20 mg, of HPP per spray. In some of the above-described embodiments, HPP is administered by a spray that can spray about 0.5 mg to about 18 mg per spray, particularly 0.5 mg to 18 mg, about 1 mg to about 16 mg per spray, particularly 1 mg to 16 mg, about 2 mg to about 14 mg per spray, particularly 2 mg to 14 mg, about 3 mg to about 12 mg per spray, particularly 3 mg to 12 mg, about 4 mg to about 10 mg per spray, particularly 4 mg to 10 mg, about 5 mg to about 9 mg per spray, particularly 5 mg to 9 mg, about 7 mg to about 8 mg per spray, particularly 7 mg to 8 mg.

[0137] In some of the above-mentioned embodiments, the dosage form is a spray. In some of the above-mentioned embodiments, the volume of the composition per spray is about 0.01 mL to about 1 mL, particularly 0.01 mL to 1 mL. In some of the above-mentioned embodiments, the volume of the composition per spray is about 0.03 mL to about 0.3 mL, particularly 0.03 mL to 0.3 mL. In some of the above-mentioned embodiments, the volume of the composition per spray is about 0.05 mL to about 0.2 mL, particularly 0.05 mL to 0.2 mL. In some of the above-mentioned embodiments, the volume of the composition per spray is about 0.07 mL to about 0.15 mL, particularly 0.07 mL to 0.15 mL. In some of the above-mentioned embodiments, the volume of the composition per spray is about 0.1 mL, particularly 0.1 mL.

[0138] In some of the above embodiments, the therapeutic strength per spray is between 0.1 mg and 50 mg of free base of HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride. In some of the above embodiments, the therapeutic strength per spray is about 1 mg to about 20 mg, particularly 1 mg to 20 mg, sometimes preferably 3 mg to 10 mg, for example 5 mg to 8 mg, or 6 mg to 7 mg of free base of HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride.

[0139] In some of the above-mentioned embodiments, the HPP of aspirin or other NSAIDs, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered by drops that can deliver about 0.05 mg to about 20 mg of HPP per drop, particularly 0.05 mg to 20 mg of HPP. In some of the above-mentioned embodiments, the HPP of aspirin or other NSAIDs, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered by drops that can deliver about 0.1 mg to about 10 mg of HPP per drop, particularly 0.1 mg to 10 mg of HPP per drop, particularly 0.2 mg to about 7 mg of HPP per drop, particularly 0.2 mg to 7 mg of HPP per drop, particularly 0.2 mg to about 1 mg of HPP per drop, particularly 0.2 mg to 1 mg of HPP per drop.

[0140] In some of the above-mentioned embodiments, the dosage form is a drop. In some of the above-mentioned embodiments, the volume of the composition per drop is about 0.01 mL to about 1 mL, particularly 0.01 mL to 1 mL. In some of the above-mentioned embodiments, the volume of the composition per drop is about 0.02 mL to about 0.3 mL, particularly 0.02 mL to 0.3 mL. In some of the above-mentioned embodiments, the volume of the composition per drop is about 0.03 mL to about 0.1 mL, particularly 0.03 mL to 0.1 mL.

[0141] In some of the above-mentioned embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered by a patch that can administer about 1 mg to about 10 g, particularly 1 mg to 10 g, of HPP per patch. In some of the above-mentioned embodiments, the HPP is administered by a patch that can administer about 50 mg to about 1 g, particularly 50 mg to 1 g, per patch. In some of the above-mentioned embodiments, the HPP is administered by a patch that can administer about 100 mg to about 500 mg, particularly 100 mg to 500 mg, per patch. In some of the above-mentioned embodiments, the HPP is administered by a patch that can administer about 200 mg to about 300 mg, particularly 200 mg to 300 mg, per patch.

[0142] In some of the above-mentioned embodiments, the dosage form is a patch. In some of the above-mentioned embodiments, the volume of the composition per patch is about 0.01 mL to about 30 mL, particularly 0.01 mL to 30 mL. In some of the above-mentioned embodiments, the volume of the composition per patch is about 0.1 mL to about 10 mL, particularly 0.1 mL to 10 mL. In some of the above-mentioned embodiments, the volume of the composition per patch is about 0.2 mL to about 2 mL, particularly 0.2 mL to 2 mL. In some of the above-mentioned embodiments, the volume of the composition per patch is about 0.5 mL to about 1 mL, particularly 0.5 mL to 1 mL.

[0143] In some of the above embodiments, the therapeutic strength per patch is 1.0 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2.0 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, g, 30mg, 35mg, 40mg, 45mg, 50mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 110mg, 120mg, 130mg, 140mg, 150mg, 160mg, 170mg , 180mg, 190mg, 200mg, 220mg, 240mg, 260mg, 280mg, 300mg, 320mg, 340mg, 360mg, 380mg, 400mg, 420mg, 440mg, 460mg, 480mg, 500mg, 200mg, 550mg , 600mg, 650mg, 700mg, 750mg, 800mg, 850mg, 900mg, 950mg, 1g, 1.1g, 1.2g, 1.3g, 1.4g, 1.5g, 1.6g, 1.7g, 1.8g, 1.9g, 2g, 2.1g, 2.2g, 2.3g, 2.4g , 2.5g, 2.6g, 2.7g, 2.8g, 2.9g, 3g, 3.1g, 3.2g, 3.3g, 3.4g, 3.5g, 3.6g, 3 .7g, 3.8g, 3.9g, 4g, 4.1g, 4.2g, 4.3g, 4.4g, 4.5g, 4.6g, 4.7g, 4.8g, 4.9 g, 5g, 5.1g, 5.2g, 5.3g, 5.4g, 5.5g, 5.6g, 5.7g, 5.8g, 5.9g, 6g, 6.1g, 6 .2g, 6.3g, 6.4g, 6.5g, 6.6g, 6.7g, 6.8g, 6.9g, 7g, 7.1g, 7.2g, 7.3g, 7.4 g, 7.5g, 7.6g, 7.7g, 7.8g, 7.9g, 8g, 8.1g, 8.2g, 8.3g, 8.4g, 8.5g, 8.6g, 8.7g, 8.8g, 8.9g, 9g, 9.1g, 9.2g, 9.3g, 9.4g, 9.5g, 9.6g, 9.7g, 9.8g, 9.9g and 10g of HPP of aspirin or other NSAID such as 2-(diethylamino)ethyl acetoxybenzoate.

[0144] In some of the above embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered once a day, twice a day, three times a day, four times a day, five times a day, or six times a day, or once a day, once every two days, once every three days, once every four days, once every five days, once every six days, or once every seven days.

[0145] In some of the above embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered once daily, twice daily, three times daily, four times daily, five times daily, six times daily, seven times daily, or eight times daily. In some embodiments, and sometimes preferred, the HPP is administered once daily, twice daily, or three times daily.

[0146] In some of the above embodiments, the HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered hourly or once every 4 to 16 hours. In some of the above embodiments, the HPP is administered hourly or once every 8 to 12 hours. In some of the above embodiments, the HPP is administered hourly or once every 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23 hours. In some of the above embodiments, HPP is administered hourly or once every 12 hours.

[0147] In some of the above embodiments, the HPP of aspirin or other NSAIDs, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, is administered from once per hour to once per day, up to once every seven days, or any frequency in between depending on the condition of the subject, preferably from once per hour to once per 24 hours, or any frequency in between, for example, once every 2 hours, once every 3 hours, once every 4 hours, once every 5 hours, once every 6 hours, once every 7 hours, once every 8 hours, once every 9 hours, once every 10 hours, once every 11 hours, once every 12 hours, once every 13 hours, once every 14 hours, once every 15 hours, once every 16 hours, once every 17 hours, once every 18 hours, once every 19 hours, once every 20 hours, once every 21 hours, once every 22 hours, once every 23 hours.

[0148] In some embodiments, and sometimes preferred, of the above embodiments, e.g., 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride aspirin or other NSAID HPP is administered once every 2 hours, once every 4 hours, once every 6 hours, once every 8 hours, once every 12 hours, once every 18 hours, or once every 24 hours. In some of the above embodiments, HPP may be administered once every 1, 2, 3, 4, 5, 6, or 7 days.

[0149] In some of the above-mentioned embodiments, the topical administration is administered for 1 day to a lifetime. In some of the above-mentioned embodiments, the topical administration is administered for 112 to 3650 consecutive or non-consecutive days. In some of the above-mentioned embodiments, the topical administration is administered for 112 to 1825 consecutive or non-consecutive days. In some of the above-mentioned embodiments, the topical administration is administered for 112 to 1095 consecutive or non-consecutive days. In some of the above-mentioned embodiments, the topical administration is administered for 112 to 730 consecutive or non-consecutive days. In some of the above-mentioned embodiments, the topical administration is administered for 112 to 365 consecutive or non-consecutive days. In some of the above-mentioned embodiments, the topical administration is administered for 112 to 224 consecutive or non-consecutive days.

[0150] In some of the above embodiments, topical administration is for at least one or more consecutive or non-consecutive days, e.g., 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, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 days, 45 days, 46 days, 47 days, 48 ​​days, 49 days, 50 days, 51 days, 52 days, 53 days, 54 days, 55 days, 56 days, 57 days, 58 days, 59 days, 60 days, 61 days, 62 days, 63 days, 64 days, 65 days, 66 days, 67 days, 68 days, 69 days, 70 days, 71 days, 72 days, 73 days, 74 days, 75 days, 76 days, 77 days, 78 days, 79 days, 80 days, 81 days, 82 days, 83 days, 84 days, 85 days, 86 days, 87 days, 88 days, 89 days, 90 days, 91 days, 92 days, 93 days, 94 days, 95 days, 96 days, 97 days, 98 days, 43 days, 44 days, 45 days, 46 days, 47 days, 48 ​​days, 49 days, 50 days, 51 days, 52 days, 53 days, 54 days, 55 days, 56 days, 57 days, 58 days, 59 days, 60 days, 61 days, 62 days, 63 days, 64 days, 65 days, 66 days, 67 days, 68 days, 69 days, 70 days, 71 days, 72 days, 73 days, 74 days, 75 days, 76 days, 77 days, 78 days, 79 days, 80 days, 81 days, 82 days, 83 days, 84 days, 85 days, 86 days, 87 days, 88 days, 89 days, 90 days, 91 days, 92 days, 93 days, 94 days, 95 days, 96 days, 97 days, 98 days, 99 days, 100 days, 101 days, 102 days, 103 days, 104 days, 105 days, 106 days, 107 days, 108 days, 109 days, 110 days, 111 days, 112 days, 113 days, 114 days, 115 days, 116 days, 117 days, 118 days, 119 days, 120 days, 121 days, 122 days, 123 days, 124 days, 125 days, 126 days, 127 days, 128 days, 129 days, 130 days, 131 days, 132 days, 133 days, 134 days, 13 5 days, 136 days, 137 days, 138 days, 139 days, 140 days, 141 days, 142 days, 143 days, 144 days, 145 days, 146 days, 147 days, 148 days, 149 days, 150 days, 155 days, 160 days, 165 days, 170 days, 175 days, 180 days, 185 days, 190 days, 195 days, 200 days, 210 days, 220 days, 230 days, 240 days, 250 days, 260 days, 270 days, 280 days, 290 days, 300 days, 320 days, 340 days, 360 days, 380 days, 400 days, 420 days,The administration is for one or more consecutive or non-consecutive days selected from 440 days, 460 days, 480 days, 500 days, 550 days, 600 days, 650 days, 700 days, 750 days, 800 days, 850 days, 900 days, 950 days, and 1000 days.

[0151] In some of the above embodiments, the topical administration is administered continuously or non-continuously for at least one year or more, e.g., for one or more consecutive or non-consecutive years selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 years, or up to the lifetime of the subject.

[0152] In some of the above-mentioned embodiments, the device of the present disclosure includes a dosage form selected from one or more of transdermal patches, creams, foams, gels, lotions, ointments, pastes, powders, shake lotions, solids, sponges, tapes, tinctures, vapors, injections, drops, rinses, sprays, and liquids. In some of the above-mentioned embodiments, the device of the present disclosure includes a dosage form selected from transdermal liquids, including one or more of transdermal drops, rinses, and sprays.

[0153] In some of the above-described embodiments, the device of the present disclosure is a device capable of administering about 0.1 mg to about 10 g, particularly 0.1 mg to 10 g, of HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, in each administration. In some of the above-described embodiments, the device of the present disclosure is a device capable of administering about 0.1 mg to about 1 g, particularly 0.1 mg to 1 g, of HPP, in each administration. In some of the above-described embodiments, the device of the present disclosure is a device capable of administering about 0.5 mg to about 500 mg, particularly 0.5 mg to 500 mg, of HPP, in each administration. In some of the above-described embodiments, the device of the present disclosure is a device capable of administering about 1 mg to about 300 mg, about 10 mg to about 300 mg, about 50 mg to about 300 mg, about 50 mg to about 200 mg, or about 200 mg to about 300 mg of HPP in each administration. In some of the above-described embodiments, the device of the present disclosure is a device capable of administering about 0.5 mg to about 30 mg, about 1 mg to about 20 mg, about 3 mg to about 10 mg, about 5 mg to about 9 mg, or about 6 mg to about 8 mg of HPP in each administration.

[0154] In some of the above-mentioned embodiments, the device is a sprayer capable of spraying about 0.1 mg to about 100 mg, particularly 0.1 mg to 100 mg, of HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, in each spray. In some of the above-mentioned embodiments, the device is a sprayer capable of spraying about 0.2 mg to about 30 mg, particularly 0.2 mg to 30 mg, of HPP in each spray. In some of the above-mentioned embodiments, the device is a sprayer capable of spraying about 0.5 mg to about 16 mg, about 1 mg to about 14 mg, about 2 mg to about 12 mg, or about 3 mg to about 10 mg of HPP in each spray. In some of the above-mentioned embodiments, the device is a sprayer capable of spraying about 4 mg to about 9 mg, about 5 mg to about 9 mg, or about 6 mg to about 9 mg of HPP in each spray. In some of the above-described embodiments, the device is a spray apparatus capable of spraying about 7 mg to about 8 mg, particularly 7 mg to 8 mg, of 2-(diethylamino)ethyl acetoxybenzoate and / or related highly penetrating prodrugs of aspirin and / or other NSAIDs or pharma- ceutically acceptable salts thereof in each spray.

[0155] In some of the above-described embodiments, the device is a spray apparatus with a nozzle that sprays about 0.1 mg to about 100 mg, particularly 0.1 mg to 100 mg, of an HPP of aspirin or other NSAID, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, each time the nozzle is pressed. In some of the above-described embodiments, the nozzle sprays about 0.2 mg to about 30 mg, about 0.5 mg to about 16 mg, about 1 mg to about 14 mg, about 2 mg to about 12 mg, 3 mg to about 11 mg, about 4 mg to about 10 mg, about 5 mg to about 9 mg, or about 6 mg to about 9 mg of HPP each time the nozzle is pressed. In some of the above-described embodiments, the device is a spray apparatus with a nozzle that sprays about 7 mg to about 8 mg, particularly 7 mg to 8 mg, of 2-(diethylamino)ethyl acetoxybenzoate and / or related highly penetrating prodrugs of aspirin and / or other NSAIDs or pharma- ceutically acceptable salts thereof with each press of the nozzle.

[0156] In some of the above-mentioned embodiments, the device is a drop capable of instilling about 0.01 mg to about 20 mg, particularly 0.01 mg to 20 mg, of HPP of aspirin or other NSAIDs, such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, in each instillation. In some of the above-mentioned embodiments, the device is a drop capable of instilling about 0.02 mg to about 18 mg, about 0.05 mg to about 16 mg, about 0.1 mg to about 14 mg, about 0.2 mg to about 12 mg, about 0.3 mg to about 10 mg, about 0.4 mg to about 9 mg, 0.5 mg to about 8 mg, or 0.7 mg to 7 mg of HPP in each instillation. In some of the above-mentioned embodiments, the device is a drop capable of instilling about 1 mg to about 6 mg, about 2 mg to about 5 mg, or about 3 mg to about 4 mg of HPP in each instillation.

[0157] In some of the above-mentioned embodiments, the device is a patch capable of administering about 0.1 mg to about 20 g, particularly 0.1 mg to 20 g, of HPP such as 2-(diethylamino)ethyl 2-acetoxybenzoate or 2-(diethylamino)ethyl acetoxybenzoate hydrochloride aspirin or other NSAID in each application. In some of the above-mentioned embodiments, the device is a patch capable of administering about 0.5 mg to about 5 g, about 1 mg to about 1000 mg, about 2 mg to about 500 mg, about 5 mg to about 400 mg, about 10 mg to about 350 mg, about 20 mg to about 300 mg, about 30 mg to about 250 mg, or about 40 mg to about 200 mg of HPP in each application. In some of the above-mentioned embodiments, the device is a patch capable of administering about 50 mg to about 200 mg, 60 mg to about 200 mg, or about 70 mg to about 200 mg of HPP in each application. In some of the above-mentioned embodiments, the device is a patch capable of administering about 100 mg to about 150 mg, particularly 100 mg to 150 mg of HPP in each application.

[0158] One aspect of the present invention is to enhance tissue penetration of NSAIDs, thereby reducing plasma drug exploration, thereby reducing side effects, and increasing tissue drug exploration, thereby enhancing drug efficacy.

[0159] In one embodiment, a composition comprising 2-(diethylamino)ethyl acetoxybenzoate hydrochloride is shown in the table below. JPEG2025504447000008.jpg45164

[0160] In one embodiment, a composition comprising 2-(diethylamino)ethyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride is shown in the table below. JPEG2025504447000009.jpg46158

[0161] In one embodiment, a composition comprising 2-(diethylamino)ethyl 2',4'-difluoro-4-hydroxyl-[1,1'-biphenyl]-3-carboxylate hydrochloride is shown in the table below. JPEG2025504447000010.jpg46158

[0162] In one embodiment, a composition comprising 2-(diethylamino)ethyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride is shown in the table below. JPEG2025504447000011.jpg53164

[0163] In one embodiment, a composition comprising 2-(diethylamino)ethyl 2-(2-hydroxybenzoyl)oxybenzoate hydrochloride is shown in the table below. JPEG2025504447000012.jpg52162

[0164] In one embodiment, a composition comprising (pyrrolidin-2-yl)methyl acetoxybenzoate hydrochloride is shown in the table below. JPEG2025504447000013.jpg46155

[0165] In one embodiment, a composition comprising (pyrrolidin-2-yl)methyl hydroxybenzoate hydrochloride is shown in the table below. JPEG2025504447000014.jpg46154

[0166] In one embodiment, a composition comprising (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride is shown in the table below. JPEG2025504447000015.jpg48159

[0167] In one embodiment, a composition comprising (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-hydroxyl-[1,1'-biphenyl]-3-carboxylate hydrochloride is shown in the table below. JPEG2025504447000016.jpg49159

[0168] In one embodiment, a composition comprising (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride is shown in the table below. JPEG2025504447000017.jpg39154

[0169] In one embodiment, a composition comprising (pyrrolidin-2-yl)methyl 2-(2-hydroxybenzoyl)oxybenzoate hydrochloride is shown in the table below. JPEG2025504447000018.jpg45141

[0170] In one embodiment, a subject will spray 5 sprays (e.g., 35 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the neck once a day, twice a day, three times a day, or four times a day until full recovery after stroke.

[0171] In one embodiment, a subject will spray 5 sprays of the medication (e.g., 35 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the neck, chest, back, abdomen, head, arms, hands, legs, feet, and other areas once a day, twice a day, three times a day, or four times a day until complete recovery from atherosclerosis.

[0172] In one embodiment, a subject will spray 5 sprays (e.g., 35 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest once a day, twice a day, three times a day, or four times a day until full recovery after a heart attack.

[0173] In one embodiment, the subject will spray 5 sprays (e.g., 35 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest once a day, twice a day, three times a day, or four times a day until complete recovery from heart failure.

[0174] In one embodiment, the subject will spray 5 sprays (e.g., 35 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest once a day, twice a day, three times a day, or four times a day until complete recovery from coronary artery disease.

[0175] In one embodiment, a subject will spray 5 sprays (e.g., 35 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest once a day, twice a day, three times a day, or four times a day until complete recovery from angina.

[0176] In one embodiment, the subject will spray 5 sprays (e.g., 35 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest once a day, twice a day, three times a day, or four times a day until complete recovery from the cardiac rhythm disorder.

[0177] In one embodiment, a subject will spray 5 sprays (e.g., 35 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest once a day, twice a day, three times a day, or four times a day until complete recovery from cardiovascular disease.

[0178] In one embodiment, a subject will spray 10 to 30 sprays (e.g., 70 mg to 210 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the neck, back, chest, legs, arms, abdomen, hands, feet, head, and other areas once a day, twice a day, three times a day, or four times a day until full recovery after stroke.

[0179] In one embodiment, the subject will spray 10 to 30 sprays (e.g., 70 mg to 210 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the neck, chest, back, abdomen, head, arms, hands, legs, feet, and other areas once a day, twice a day, three times a day, or four times a day until complete recovery from atherosclerosis.

[0180] In one embodiment, a subject will spray 10 to 30 sprays (e.g., 70 mg to 210 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest, neck, back, abdomen, and other areas once a day, twice a day, three times a day, or four times a day until a complete recovery after the heart attack.

[0181] In one embodiment, the subject will spray 10 to 30 sprays (e.g., 70 mg to 210 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest, neck, back, abdomen, legs, arms, and other areas once a day, twice a day, three times a day, or four times a day until complete recovery from heart failure.

[0182] In one embodiment, the subject will spray 10 to 30 sprays (e.g., 70 mg to 210 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest, neck, back, abdomen, legs, arms, and other areas once a day, twice a day, three times a day, or four times a day until complete recovery from coronary artery disease.

[0183] In one embodiment, the subject will spray 10 to 30 sprays (e.g., 70 mg to 210 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest, neck, back, abdomen, legs, arms, and other areas once a day, twice a day, three times a day, or four times a day until complete recovery from angina.

[0184] In one embodiment, the subject will spray 10 to 30 sprays (e.g., 70 mg to 210 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest, neck, back, abdomen, legs, arms, and other areas once a day, twice a day, three times a day, or four times a day until complete recovery from the cardiac rhythm disorder.

[0185] In one embodiment, the subject will spray 10 to 30 sprays (e.g., 70 mg to 210 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol) onto the skin around the chest, neck, back, abdomen, legs, arms, and other areas once a day, twice a day, three times a day, or four times a day until complete recovery from the cardiovascular disease. EXAMPLES

[0186] The following non-limiting examples further illustrate certain aspects of the present invention.

[0187] Example 1 Preparation of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride (aspirin HPP) Acetoxybenzoyl chloride (20 g) was dissolved in ethyl acetate (100 ml). The mixture was cooled to 0° C. Diethylaminoethanol (14 g) was added to the reaction mixture. The mixture was stirred at room temperature for 3 hours and then washed with water (5×30 ml). 3N HCl in ethanol (30 ml) was added and the solid was collected, washed with ethyl acetate (5×50 ml) and dried in a vacuum oven at 40° C. (90% yield).

[0188] Example 2 Preparation of 2-(diethylamino)ethyl hydroxybenzoate hydrochloride (HPP of salicylic acid) 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride (20 g) was dissolved in water (100 ml) and 3N HCl (10 ml). The mixture was stirred at room temperature overnight. Ethyl acetate (300 ml) was added to the mixture, the pH of the mixture was adjusted to 8 with solid NaHCO3, the ethyl acetate layer was collected, washed with water (3 times), 1N HCl in ethyl acetate (100 ml) was added, the solid was collected, washed with ethyl acetate (5 x 20 ml), and dried in a vacuum oven at 40°C (72% yield).

[0189] Example 3 Preparation of 2-(diethylamino)ethyl 5-(2,4-difluorophenyl)acetoxybenzoate hydrochloride (acetyldiflunisal HPP) 5-(2,4-Difluorophenyl)acetoxybenzoyl chloride (2',4'-difluoro-4-hydroxy-[1,1'-biphenyl]-3-carbonyl chloride) (31 g) was dissolved in ethyl acetate (100 ml). The mixture was cooled to 0° C. Diethylaminoethanol (14 g) was added to the reaction mixture. The mixture was stirred at room temperature for 3 hours. The ethyl acetate mixture was washed with water (5×30 ml). 3N HCl in ethanol (30 ml) was added and the solid was collected, washed with ethyl acetate (5×50 ml) and dried in a vacuum oven at 40° C. (88% yield).

[0190] Example 4 Preparation of 2-(diethylamino)ethyl 2',4'-difluoro-4-hydroxyl-[1,1'-biphenyl]-3-carboxylate hydrochloride (Diflunisal HPP) 2',4'-Difluoro-4-hydroxy-[1,1'-biphenyl]-3-carboxylate hydrochloride (20 g) was dissolved in water (100 ml) and 3N HCl (10 ml). The mixture was stirred at room temperature overnight. Ethyl acetate (300 ml) was added to the mixture, the pH of the mixture was adjusted to 8 with solid NaHCO3, the ethyl acetate layer was collected, washed with water (3 times), 1N HCl in ethyl acetate (100 ml) was added, the solid was collected, washed with ethyl acetate (5 x 20 ml), and dried in a vacuum oven at 40°C (80% yield).

[0191] Example 5 Preparation of (pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride (aspirin HPP) Acetylsalicylic acid (18 g) and N-Boc-L-prolinol (tert-butoxycarbonyl-2-pyrrolidinemethanol, 20.1 g) were placed in a 1 L round bottom flask and acetone (200 ml) was added to the mixture. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI, 19.2 g), 4-dimethylaminopyridine (DMAP, 5 g), and 1-hydroxybenzotriazole (HOBt, 15 g) were added to the solution. The mixture was stirred overnight at room temperature. The solution was evaporated to near dryness. Ethyl acetate (500 ml) was added to the mixture. The solution was washed with water (2 x 200 ml), 20% citric acid (50 g in 250 ml water) (2 x 250 ml), and water (3 x 300 ml). The solution was dried over sodium sulfate. The sodium sulfate was removed by filtration, the filtrate was washed with ethyl acetate (3 x 50 ml) and the ethyl acetate solution was evaporated to dryness. 3N HCl in ethyl acetate (50 ml) was added and the mixture was stirred for 3 hours. The solid was collected, washed with ethyl acetate (5 x 50 ml) and dried in a vacuum oven at 40°C (85% yield).

[0192] Example 6 Preparation of (pyrrolidin-2-yl)methyl hydroxybenzoate hydrochloride (Pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride (20 g) was dissolved in water (100 ml) and 3N HCl (10 ml). The mixture was stirred at room temperature overnight. Ethyl acetate (300 ml) was added to the mixture, the pH of the mixture was adjusted to 8 with solid NaHCO3, the ethyl acetate layer was collected, washed with water (3 times), 1N HCl in ethyl acetate (100 ml) was added, the solid was collected, washed with ethyl acetate (5 x 20 ml), and dried in a vacuum oven at 40°C (80% yield).

[0193] Example 7 Preparation of (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride 2',4'-Difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylic acid (31 g) and N-Boc-L-prolinol (tert-butoxycarbonyl-2-pyrrolidinemethanol, 20.1 g) were placed in a 1 L round bottom flask and acetone (200 ml) was added to the mixture. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI, 19.2 g), 4-dimethylaminopyridine (DMAP, 5 g), and 1-hydroxybenzotriazole (HOBt, 15 g) were added to the solution. The mixture was stirred overnight at room temperature. The solution was evaporated to near dryness. Ethyl acetate (500 ml) was added to the mixture. The solution was washed with water (2 x 200 ml), 20% citric acid (50 g in 250 ml water) (2 x 250 ml), and water (3 x 300 ml). The solution was dried over sodium sulfate. The sodium sulfate was removed by filtration, the filtrate was washed with ethyl acetate (3 x 50 ml) and the ethyl acetate solution was evaporated to dryness. 3N HCl in ethyl acetate (50 ml) was added and the mixture was stirred for 3 hours. The solid was collected, washed with ethyl acetate (5 x 50 ml) and dried in a vacuum oven at 40°C (85% yield).

[0194] Example 8 Preparation of (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-hydroxyl-[1,1'-biphenyl]-3-carboxylate hydrochloride (Pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride (20 g) was dissolved in water (100 ml) and 3N HCl (10 ml). The mixture was stirred at room temperature overnight. Ethyl acetate (300 ml) was added to the mixture, the pH of the mixture was adjusted to 8 with solid NaHCO3, the ethyl acetate layer was collected, washed with water (3 x 100 ml), 1N HCl in ethyl acetate (100 ml) was added, the solid was collected, washed with ethyl acetate (5 x 20 ml), and dried in a vacuum oven at 40°C (80% yield).

[0195] Example 9 Preparation of 2-(diethylamino)ethyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride (prodrug of acetylsalsalate) 2-(2-acetoxybenzoyl)oxybenzoyl chloride (28 g) was dissolved in ethyl acetate (100 ml). The mixture was cooled to 0° C. Diethylaminoethanol (14 g) was added to the reaction mixture. The mixture was stirred at room temperature for 3 hours. The ethyl acetate mixture was washed with water (5×30 ml). 3N HCl in ethanol (30 ml) was added and the solid was collected, washed with ethyl acetate (5×50 ml) and dried in a vacuum oven at 40° C. (91% yield).

[0196] Example 10 Preparation of 2-(diethylamino)ethyl 2-(2-hydroxybenzoyl)oxybenzoate hydrochloride (prodrug of salsalate) 2-(Diethylamino)ethyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride (20 g) was dissolved in water (100 ml) and 3N HCl (10 ml). The mixture was stirred at room temperature overnight. Ethyl acetate (300 ml) was added to the mixture, the pH of the mixture was adjusted to 8 with solid NaHCO3, the ethyl acetate layer was collected, washed with water (3 x 100 ml), 1N HCl in ethyl acetate (100 ml) was added, the solid was collected, washed with ethyl acetate (5 x 20 ml), and dried in a vacuum oven at 40°C (68% yield).

[0197] Example 11 Preparation of (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride 2-(2-acetoxybenzoyl)oxybenzoic acid (28 g) and N-Boc-L-prolinol (tert-butoxycarbonyl-2-pyrrolidinemethanol, 20.1 g) were placed in a 1 L round bottom flask and acetone (200 ml) was added to the mixture. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI, 19.2 g), 4-dimethylaminopyridine (DMAP, 5 g), and 1-hydroxybenzotriazole (HOBt, 15 g) were added to the solution. The mixture was stirred overnight at room temperature. The solution was evaporated to near dryness. Ethyl acetate (500 ml) was added to the mixture. The solution was washed with water (2 x 200 ml), 20% citric acid (R0089, 50 g in 250 ml water) (2 x 250 ml), and water (3 x 300 ml). The solution was dried over sodium sulfate. The sodium sulfate was removed by filtration, the filtrate was washed with ethyl acetate (3 x 50 ml) and the ethyl acetate solution was evaporated to dryness. 3N HCl in ethyl acetate (50 ml) was added and the mixture was stirred for 3 hours. The solid was collected, washed with ethyl acetate (5 x 50 ml) and dried in a vacuum oven at 40°C (85% yield).

[0198] Example 12 Preparation of (pyrrolidin-2-yl)methyl 2-(2-hydroxybenzoyl)oxybenzoate hydrochloride (Pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride (20 g) was dissolved in water (100 ml) and 3N HCl (10 ml). The mixture was stirred at room temperature overnight. Ethyl acetate (300 ml) was added to the mixture, the pH of the mixture was adjusted to 8 with solid NaHCO3, the ethyl acetate layer was collected, washed with water (3 x 100 ml), 1N HCl in ethyl acetate (100 ml) was added, the solid was collected, washed with ethyl acetate (5 x 20 ml), and dried in a vacuum oven at 40°C (80% yield).

[0199] Example 13 Skin permeability of HPP 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride (Compound-1), (Pyrrolidin-2-yl)methyl acetoxybenzoate hydrochloride (Compound-2), Aspirin (Compound-3), 2-(Diethylamino)ethyl hydroxybenzoate hydrochloride (Compound-4), 2(Pyrrolidin-2-yl)methyl hydroxybenzoate hydrochloride (Compound-5), Salicylic acid (Compound-6), 2-(Diethylamino)ethyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate 2-(Diethylamino)ethyl 2',4'-difluoro-4-hydroxyl-[1,1'-biphenyl]-3-carboxylate hydrochloride (Compound-7), (Pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride (Compound-8), 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylic acid (Compound-9), 2-(Diethylamino)ethyl 2',4'-difluoro-4-hydroxyl-[1,1'-biphenyl]-3-carboxylate hydrochloride (Compound-10), (Pyrrolidin-2-yl)methyl 2',4'-difluoro-4- Hydroxyl-[1,1'-biphenyl]-3-carboxylate hydrochloride (Compound-11), 2',4'-difluoro-4-hydroxyl-[1,1'-biphenyl]-3-carboxylic acid (Diflunisal, Compound-12), 2-(Diethylamino)ethyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride (Compound-13), (Pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride (Compound-14), 2-(2-acetoxybenzoyl)oxybenzoic acid (Acetyl) The permeability of 2-(diethylamino)ethyl 2-(2-hydroxybenzoyl)oxybenzoate hydrochloride (compound-16), (pyrrolidin-2-yl)methyl 2-(2-hydroxybenzoyl)oxybenzoate hydrochloride (compound-17), and 2-(2-hydroxybenzoyl)oxybenzoic acid (salsalate, compound-18) through rabbit skin was measured in vitro by using modified Franz cells isolated from rabbit dorsal skin tissue (300 μm to 350 μm thick). The receptor solution, consisting of 10 ml of pure water, is shown in Table 1.These results suggest that transport units play a crucial role in drug passage across membrane and skin barriers.

[0200] Table 1. Cumulative doses of anti-inflammatory drugs over an 8-hour period JPEG2025504447000019.jpg117145

[0201] Example 14 Efficacy of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in rats with acute ischemia induced by middle cerebral artery occlusion (MCAO) The aim of this study was to investigate the therapeutic efficacy of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride on cerebral ischemic injury and associated neurological deficits induced by transient MCAO in rats.

[0202] Male SD rats (258g-280g) were used. Rats were selected for inclusion based on acceptable clinical condition and body weight. Animals were randomly assigned to seven groups (25 rats per group for transdermal 2-(diethylamino)ethyl acetoxybenzoate hydrochloride treatment, oral aspirin treatment, MCAO surgery including MCI-186 / Edaravone (intravenous) treatment, and vehicle group, and 8 rats per group for sham operation group). Rats were kept in the animal facility for one week before all procedures. Animal identification numbers were labeled on the cage tags as well as on the tails. See the experimental design shown in Table 2.

[0203] 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride, aspirin, edaravone, and vehicle were administered 1 hour after MCAO surgery (day 0) and on days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14.

[0204] Table 2. Experimental design JPEG2025504447000020.jpg66159

[0205] Prior to MCAO, animals were fasted overnight but allowed free access to water. Under anesthesia, a midline incision was made in the neck to expose the right common carotid artery (CCA), internal carotid artery (ICA), and external carotid artery (ECA). A commercially available monofilament (with silicone coating) was used as an occlusion plug and inserted through the CCA. The occlusion plug was advanced into the CCA 18±0.5 mm beyond the carotid bifurcation. Light resistance indicated that the occlusion plug was adequately lodged within the anterior cerebral artery, thus blocking blood flow to the middle cerebral artery (MCA). After 1 hour, reperfusion was allowed by fully withdrawing the monofilament. Body temperature was maintained at approximately 36.5°C during the surgical procedure using a heating pad. At 2 hours after occlusion, clinical signs were examined by an observer blinded to the treatment groups to confirm neurological deficits. Neurological deficits (see Table 3) were assessed daily thereafter until the 14th day. 14 days after MCAO, the animals were euthanized and the brains were cut into 5 coronal sections (2 mm thick by using a rat brain matrix). Fresh brain sections were stained with 2% triphenyltetrazolium chloride (TTC) solution at 37°C, and then fixed with 4% paraformaldehyde. Photographs of all sections were taken with a digital camera. These digital photographs were transferred to a computer. The infarct volume per brain was obtained after blindly measuring the infarct area (%) for each section using Image-Pro Plus software. Infarct area (%) = (contralateral hemisphere area - ipsilateral non-infarct area) / contralateral hemisphere area, where the relevant swelling and atrophy were corrected by this formula.

[0206] Table 3. Neurological signs scores JPEG2025504447000021.jpg60148

[0207] For the analysis of infarct size, One-Way ANOVA followed by Dunnett's multiple comparison test was used to compare the differences between the treatment and vehicle groups. A t-test was used to compare the differences between the positive control and vehicle groups. P<0.05 was considered to indicate a significant difference between groups. Data were expressed as mean±SEM. For the analysis of body weight, a t-test was used to compare the differences between groups at various days after surgery.

[0208] All animals were in good general condition before the study. During the experiment, mortality was 5 of 25 (20%) in the vehicle group, 6 of 25 (24%) in the edaravone-treated group, 3 of 25 (12%) in the oral aspirin group, 1 of 25 (4%) in the low-dose transdermal 2-(diethylamino)ethyl acetoxybenzoate hydrochloride group, 0 of 25 in the medium- and high-dose transdermal 2-(diethylamino)ethyl acetoxybenzoate hydrochloride groups, and 0 of 8 (0%) in the sham-operated group. Most of the animal deaths occurred between 48 hours (6 rats) and 72 hours after the MCAO operation. All other MCAO animals lost weight from the first to the fourth day after the operation and then maintained a relatively stable health condition until euthanasia. Animals in the sham-operated group lost weight only during the first 2 days after the sham operation and then gradually returned to normal levels.

[0209] Several animals with obvious bleeding during surgery were abandoned. After the final stage of the surgery (removal of the occluding plug), the animals were closely monitored until they had fully regained consciousness.

[0210] Rats treated with transdermal 2-(diethylamino)ethyl acetoxybenzoate hydrochloride showed smaller infarct volumes compared to intravenous edaravone (38.07%±2.031), oral aspirin (36.27%±2.123), and vehicle groups (40.53%±2.378), expressed as mean±SEM: low dose group (32.07%±2.061, P<0.05), medium dose group (27.11%±1.658, P<0.01), and high dose group (25.15%±2.001, P<0.01). The differences of all low, medium, and high dose groups against the vehicle group were statistically significant (P<0.05 or P<0.01). In contrast, aspirin- and MCI-186 / Edaravone-treated rats showed little reduction in infarct volume. Rats in the sham group showed unavoidable measurement variation (2.85% ± 0.779). Most of the infarcts in the vehicle-, aspirin-, and MCI-186 / Edaravone-treated groups were typically involved in the striatum and frontoparietal cortex supplied by the middle cerebral artery, whereas the infarct size was smaller and the striatum and corresponding cortex were less involved in the low-, medium-, and high-dose groups. Detailed analysis of infarct volume is shown in Figure 1.

[0211] The body weight of MACO rats dropped dramatically during the first 4 days after surgery and then remained relatively stable. Meanwhile, the body weights of the low (P<0.05), medium (P<0.01), and high (P<0.01) dose groups of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride increased from day 5 to day 14 and were significantly different from the vehicle group by t-test. Detailed information on the body weight assessment is shown in Figure 2.

[0212] Ischemic injury caused clinical signs of motor dysfunction on the left side. Significant differences were observed in the neurological deficit scores of the low, medium, and high dose groups of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride compared to the vehicle group. Detailed information on the assessment of neurological deficit scores is shown in Figure 3.

[0213] This study investigated the therapeutic efficacy of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride against cerebral ischemia in a rat MCAO model.Compared with the vehicle group, the low-, medium-, and high-dose groups of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride showed good neuroprotective effects against cerebral ischemic injury.

[0214] Example 15 Efficacy of 2-(diethylamino)ethyl hydroxybenzoate hydrochloride in rats with acute ischemia induced by middle cerebral artery occlusion (MCAO) This study was conducted to investigate the therapeutic efficacy of 2-(diethylamino)ethyl hydroxybenzoate on cerebral ischemic injury and associated neurological deficits induced by transient MCAO in rats.

[0215] Male SD rats (258g-275g) were used. Rats were selected for inclusion based on acceptable clinical condition and body weight. Animals were randomly assigned to seven groups (25 rats per group for transdermal 2-(diethylamino)ethyl hydroxybenzoate hydrochloride treatment, oral aspirin treatment, MCAO surgery including MCI-186 / Edaravone (intravenous) treatment, and vehicle group, and 8 rats per group for sham operation group). Rats were kept in the animal facility for one week before all procedures. Animal identification numbers were labeled on the cage tags as well as on the tails. See the experimental design shown in Table 4.

[0216] 2-(Diethylamino)ethyl hydroxybenzoate hydrochloride, salicylic acid, edaravone, and vehicle were administered 1 hour after MCAO surgery (day 0) and on days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14.

[0217] Table 4. Experimental design JPEG2025504447000022.jpg66159

[0218] Prior to MCAO, animals were fasted overnight but allowed free access to water. Under anesthesia, a midline incision was made in the neck to expose the right common carotid artery (CCA), internal carotid artery (ICA), and external carotid artery (ECA). A commercially available monofilament (with silicone coating) was used as an occlusion plug and inserted through the CCA. The occlusion plug was advanced into the CCA 18±0.5 mm beyond the carotid bifurcation. Light resistance indicated that the occlusion plug was adequately lodged within the anterior cerebral artery, thus blocking blood flow to the middle cerebral artery (MCA). After 1 hour, reperfusion was allowed by fully withdrawing the monofilament. Body temperature was maintained at approximately 36.5°C during the surgical procedure using a heating pad. At 2 hours after occlusion, clinical signs were examined by an observer blinded to the treatment groups to confirm neurological deficits. Neurological deficits (see Table 3) were assessed daily thereafter until the 14th day. 14 days after MCAO, the animals were euthanized and the brains were cut into 5 coronal sections (2 mm thick by using a rat brain matrix). Fresh brain sections were stained with 2% triphenyltetrazolium chloride (TTC) solution at 37°C, and then fixed with 4% paraformaldehyde. Photographs of all sections were taken with a digital camera. These digital photographs were transferred to a computer. The infarct volume per brain was obtained after blindly measuring the infarct area (%) for each section using Image-Pro Plus software. Infarct area (%) = (contralateral hemisphere area - ipsilateral non-infarct area) / contralateral hemisphere area, where the relevant swelling and atrophy were corrected by this formula.

[0219] For the analysis of infarct size, One-Way ANOVA followed by Dunnett's multiple comparison test was used to compare the differences between the treatment and vehicle groups. A t-test was used to compare the differences between the positive control and vehicle groups. P<0.05 was considered to indicate a significant difference between groups. Data were expressed as mean±SEM. For the analysis of body weight, a t-test was used to compare the differences between groups at various days after surgery.

[0220] All animals were in good general condition before the study. During the experiment, mortality was 5 of 25 (20%) in the vehicle group, 5 of 25 (20%) in the edaravone-treated group, 4 of 25 (16%) in the oral salicylic acid group, 3 of 25 (12%) in the low-dose transdermal 2-(diethylamino)ethyl acetoxybenzoate hydrochloride group, 2 of 25 (8%) in the medium-dose transdermal 2-(diethylamino)ethyl hydroxybenzoate hydrochloride group, 1 of 25 (4%) in the high-dose transdermal 2-(diethylamino)ethyl hydroxybenzoate hydrochloride group, and 0 of 8 (0%) in the sham-operated group. Most of the animal deaths occurred between 48 hours (6 rats) and 72 hours after the MCAO operation. All other MCAO animals lost weight between days 1 and 4 after surgery and then maintained a relatively stable health condition until euthanasia. Animals in the sham-operated group lost weight only during the first 2 days after sham surgery, after which the weight gradually returned to normal levels.

[0221] Several animals with obvious bleeding during surgery were abandoned. After the final stage of the surgery (removal of the occluding plug), the animals were closely monitored until they had fully regained consciousness.

[0222] Rats treated with transdermal 2-(diethylamino)ethyl hydroxybenzoate hydrochloride showed smaller infarct volumes compared to the intravenous edaravone (39.94% ± 2.256), oral salicylic acid (37.47% ± 2.856), and vehicle groups (40.95% ± 2.567), expressed as mean ± SEM: low dose group (37.01% ± 2.053, P < 0.08), medium dose group (35.77% ± 1.875, P < 0.07), and high dose group (34.56% ± 2.563, P < 0.05). The difference of the high dose group against the vehicle group was statistically significant (P < 0.05). In contrast, salicylic acid- and MCI-186 / edaravone-treated rats showed little reduction in infarct volume. Rats in the sham group showed unavoidable measurement variation (2.12% ± 0.853). Most of the infarcts in the vehicle-, salicylic acid-, and MCI-186 / edaravone-treated groups were typically involved in the striatum and frontoparietal cortex supplied by the middle cerebral artery, whereas the infarct size was smaller and the striatum and corresponding cortex were less involved in the low-, medium-, and high-dose groups. Detailed analysis of infarct volume is shown in Figure 4.

[0223] Body weight in MACO rats dropped dramatically during the first 4 days after surgery and then remained relatively stable. Meanwhile, body weights in the low (P<0.11), medium (P<0.08), and high (P<0.05) dose groups of 2-(diethylamino)ethyl hydroxybenzoate hydrochloride (a prodrug of salicylic acid) increased from day 5 to day 14 and were significantly different from the vehicle group by t-test. Detailed information on body weight assessment is shown in Figure 5.

[0224] Ischemic injury caused clinical signs of motor dysfunction on the left side. The neurological deficit scores of the high dose 2-(diethylamino)ethyl hydroxybenzoate hydrochloride group compared to the vehicle group showed significant differences. Detailed information on the assessment of the neurological deficit scores is shown in Figure 6.

[0225] This study investigated the therapeutic efficacy of 2-(diethylamino)ethyl hydroxybenzoate hydrochloride against cerebral ischemia in a rat MCAO model. Compared with the vehicle group, the high-dose group of 2-(diethylamino)ethyl hydroxybenzoate hydrochloride showed good neuroprotective effects against cerebral ischemic injury.

[0226] Example 16 Efficacy of (pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride in rats with acute ischemia induced by middle cerebral artery occlusion (MCAO) The aim of this study was to investigate the therapeutic efficacy of 2-(diethylamino)ethyl acetoxybenzoate on cerebral ischemic injury and associated neurological deficits induced by transient MCAO in rats.

[0227] Male SD rats (255g-283g) were used. Rats were selected for inclusion based on acceptable clinical condition and body weight. Animals were randomly assigned to seven groups (25 rats per group for transdermal (pyrrolidin-2-yl) methylacetoxybenzoate hydrochloride treatment, oral aspirin treatment, MCAO surgery including MCI-186 / Edaravone (intravenous) treatment, and vehicle group, and 8 rats per group for sham operation group). Rats were kept in the animal facility for one week before all procedures. Animal identification numbers were labeled on the cage tags as well as on the tails. See the experimental design shown in Table 5.

[0228] (Pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride, aspirin, edaravone, and vehicle were administered 1 hour after MCAO surgery (day 0) and on days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14.

[0229] Table 5. Experimental design JPEG2025504447000023.jpg66158

[0230] Prior to MCAO, animals were fasted overnight but allowed free access to water. Under anesthesia, a midline incision was made in the neck to expose the right common carotid artery (CCA), internal carotid artery (ICA), and external carotid artery (ECA). A commercially available monofilament (with silicone coating) was used as an occlusion plug and inserted through the CCA. The occlusion plug was advanced into the CCA 18±0.5 mm beyond the carotid bifurcation. Light resistance indicated that the occlusion plug was adequately lodged within the anterior cerebral artery, thus blocking blood flow to the middle cerebral artery (MCA). After 1 hour, reperfusion was allowed by fully withdrawing the monofilament. Body temperature was maintained at approximately 36.5°C during the surgical procedure using a heating pad. At 2 hours after occlusion, clinical signs were examined by an observer blinded to the treatment groups to confirm neurological deficits. Neurological deficits (see Table 3) were assessed daily thereafter until the 14th day. 14 days after MCAO, the animals were euthanized and the brains were cut into 5 coronal sections (2 mm thick by using a rat brain matrix). Fresh brain sections were stained with 2% triphenyltetrazolium chloride (TTC) solution at 37°C, and then fixed with 4% paraformaldehyde. Photographs of all sections were taken with a digital camera. These digital photographs were transferred to a computer. The infarct volume per brain was obtained after blindly measuring the infarct area (%) for each section using Image-Pro Plus software. Infarct area (%) = (contralateral hemisphere area - ipsilateral non-infarct area) / contralateral hemisphere area, where the relevant swelling and atrophy were corrected by this formula.

[0231] For the analysis of infarct size, One-Way ANOVA followed by Dunnett's multiple comparison test was used to compare the differences between the treatment and vehicle groups. A t-test was used to compare the differences between the positive control and vehicle groups. P<0.05 was considered to indicate a significant difference between groups. Data were expressed as mean±SEM. For the analysis of body weight, a t-test was used to compare the differences between groups at various days after surgery.

[0232] All animals were in good general condition before the study. During the experiment, mortality was 5 of 25 (20%) in the vehicle group, 5 of 25 (20%) in the edaravone-treated group, 3 of 25 (12%) in the oral aspirin group, 1 of 25 (4%) in the low-dose transdermal (pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride group, 0 of 25 in the medium-dose and high-dose transdermal (pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride groups, and 0 of 8 (0%) in the sham-operated group. Most of the animal deaths occurred between 48 hours (6 rats) and 72 hours after the MCAO operation. All other MCAO animals lost weight from the first to the fourth day after the operation and then maintained a relatively stable health condition until euthanasia. Animals in the sham-operated group lost weight only during the first 2 days after the sham operation and then gradually returned to normal levels.

[0233] Several animals with obvious bleeding during surgery were abandoned. After the final stage of the surgery (removal of the occluding plug), the animals were closely monitored until they had fully regained consciousness.

[0234] Rats treated with transdermal (pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride showed smaller infarct volumes compared to intravenous edaravone (39.27%±2.513), oral aspirin (37.99%±1.985), and vehicle groups (41.78%±2.179), expressed as mean±SEM: low dose group (33.11%±2.212, P<0.05), medium dose group (27.03%±2.123, P<0.01), and high dose group (24.85%±2.376, P<0.01). The differences of all low, medium, and high dose groups against the vehicle group were statistically significant (P<0.05 or P<0.01). In contrast, aspirin- and MCI-186 / Edaravone-treated rats showed little reduction in infarct volume. Rats in the sham group showed unavoidable measurement variation (3.12%±1.279). Most of the infarcts in the vehicle-, aspirin-, and MCI-186 / Edaravone-treated groups were typically involved in the striatum and frontoparietal cortex supplied by the middle cerebral artery, whereas the infarct size was smaller and the striatum and corresponding cortex were less involved in the low-, medium-, and high-dose groups. Detailed analysis of infarct volume is shown in FIG. 7.

[0235] The body weight of MACO rats dropped dramatically during the first 4 days after surgery and then remained relatively stable. Meanwhile, the body weights of the low (P<0.05), medium (P<0.01), and high (P<0.01) dose groups of (pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride increased from day 5 to day 14, and were significantly different from the vehicle group by t-test. Detailed information on the body weight assessment is shown in Figure 8.

[0236] Ischemic injury caused clinical signs of motor dysfunction on the left side. The neurological deficit scores of the low, medium and high dose groups of (pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride compared with the vehicle group show significant differences. Detailed information on the assessment of the neurological deficit scores is shown in Figure 9.

[0237] This study investigated the therapeutic efficacy of (pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride against cerebral ischemia in a rat MCAO model. Compared with the vehicle group, the low-, medium-, and high-dose groups of (pyrrolidin-2-yl)methylacetoxybenzoate hydrochloride showed good neuroprotective effects against cerebral ischemic injury.

[0238] Example 17 Efficacy of (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride in rats with acute ischemia induced by middle cerebral artery occlusion (MCAO) The aim of this study was to investigate the therapeutic efficacy of (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride on cerebral ischemic injury and associated neurological deficits induced by transient MCAO in rats.

[0239] Male SD rats (257g-281g) were used. Rats were selected for inclusion based on acceptable clinical condition and body weight. Animals were randomly assigned to seven groups (25 rats per group for transdermal (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride treatment, acetyldiflunisal treatment, MCAO surgery including MCI-186 / Edaravone (intravenous) treatment, and vehicle group, and 8 rats per group for sham operation group). Rats were kept in the animal facility for one week before all procedures. Animal identification numbers were labeled on the cage tags as well as on the tails. See the experimental design shown in Table 6.

[0240] (Pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride, 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylic acid (acetyldiflunisal), edaravone, and vehicle were administered 1 hour after MCAO surgery (day 0) and on days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14.

[0241] Table 6. Experimental design JPEG2025504447000024.jpg66162

[0242] Prior to MCAO, animals were fasted overnight but allowed free access to water. Under anesthesia, a midline incision was made in the neck to expose the right common carotid artery (CCA), internal carotid artery (ICA), and external carotid artery (ECA). A commercially available monofilament (with silicone coating) was used as an occluder and inserted through the CCA. The occluder was advanced into the CCA 18±0.5 mm beyond the carotid bifurcation. Light resistance indicated that the occluder was adequately lodged within the anterior cerebral artery, thus blocking blood flow to the middle cerebral artery (MCA). After 1 hour, reperfusion was allowed by fully withdrawing the monofilament. Body temperature was maintained at approximately 36.5°C during the surgical procedure using a heating pad. At 2 hours after occlusion, clinical signs were examined by an observer blinded to the treatment groups to confirm neurological deficits. Neurological deficits (see Table 3) were assessed daily thereafter until the 14th day. 14 days after MCAO, the animals were euthanized and the brains were cut into 5 coronal sections (2 mm thick by using a rat brain matrix). Fresh brain sections were stained with 2% triphenyltetrazolium chloride (TTC) solution at 37°C, and then fixed with 4% paraformaldehyde. Photographs of all sections were taken with a digital camera. These digital photographs were transferred to a computer. The infarct volume per brain was obtained after blindly measuring the infarct area (%) for each section using Image-Pro Plus software. Infarct area (%) = (contralateral hemisphere area - ipsilateral non-infarct area) / contralateral hemisphere area, where the relevant swelling and atrophy were corrected by this formula.

[0243] For the analysis of infarct size, One-Way ANOVA followed by Dunnett's multiple comparison test was used to compare the differences between the treatment and vehicle groups. A t-test was used to compare the differences between the positive control and vehicle groups. P<0.05 was considered to indicate a significant difference between groups. Data were expressed as mean±SEM. For the analysis of body weight, a t-test was used to compare the differences between groups at various days after surgery.

[0244] All animals were in good general condition before the study. During the experiment, mortality was 4 of 25 (16%) in the vehicle group, 5 of 25 (20%) in the edaravone treatment group, 3 of 25 (12%) in the oral acetyldiflunisal group, 2 of 25 (8%) in the low dose transdermal (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride group, 1 of 25 (4%) in the medium and high dose transdermal (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride groups, and 0 of 8 (0%) in the sham operation group. Most of the animal deaths occurred between 48 hours (6 rats) and 72 hours after the MCAO operation. All other MCAO animals lost weight between days 1 and 4 after surgery and then maintained a relatively stable health status until euthanasia. Animals in the sham-operated group lost weight only during the first 2 days after sham surgery and then gradually returned to normal levels.

[0245] Several animals with obvious bleeding during surgery were abandoned. After the final stage of the surgery (removal of the occluding plug), the animals were closely monitored until they had fully regained consciousness.

[0246] Rats treated with transdermal (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride showed smaller infarct volumes compared to intravenous edaravone (40.35% ± 2.078), oral acetyldiflunisal (38.95% ± 2.176), and vehicle groups (41.08% ± 1.982), expressed as mean ± SEM: low dose group (37.52% ± 2.789, P < 0.12), medium dose group (35.02% ± 2.827, P < 0.06), and high dose group (33.53% ± 1.986, P < 0.05). The difference of the high dose group against the vehicle group was statistically significant (P < 0.05). In contrast, acetyl diflunisal and MCI-186 / edaravone treated rats showed little reduction in infarct volume. Rats in the sham group showed unavoidable measurement variation (1.98% ± 0.751). Most of the infarcts in the vehicle, acetyl diflunisal and MCI-186 / edaravone treated groups were typically involved in the striatum and frontoparietal cortex supplied by the middle cerebral artery, whereas the infarct size was smaller and the striatum and corresponding cortex were less involved in the low, medium and high dose groups. Detailed analysis of infarct volume is shown in Figure 10.

[0247] The body weight of MACO rats dropped dramatically during the first 4 days after surgery and then remained relatively stable. Meanwhile, the body weights of the low (P<0.10), medium (P<0.08), and high (P<0.05) dose groups of (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride increased from day 5 to day 14 and were significantly different from the vehicle group by t-test. Detailed information on the body weight assessment is shown in Figure 11.

[0248] Ischemic damage caused clinical signs of motor dysfunction on the left side. The neurological deficit scores of the high dose (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride group compared to the vehicle group show significant differences. Detailed information on the assessment of neurological deficit scores is shown in Figure 12.

[0249] This study investigated the therapeutic efficacy of (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride against cerebral ischemia in a rat MCAO model. Compared with the vehicle group, the high-dose group of (pyrrolidin-2-yl)methyl 2',4'-difluoro-4-acetoxy-[1,1'-biphenyl]-3-carboxylate hydrochloride showed good neuroprotective effects against cerebral ischemic injury.

[0250] Example 18 Efficacy of (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride in rats with acute ischemia induced by middle cerebral artery occlusion (MCAO) The aim of this study was to investigate the therapeutic efficacy of (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride on cerebral ischemic injury and associated neurological deficits induced by transient MCAO in rats.

[0251] Male SD rats (255g-280g) were used. Rats were selected for inclusion based on acceptable clinical condition and body weight. Animals were randomly assigned to seven groups (25 rats per group for transdermal (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride treatment, acetylsalsalate treatment, MCAO surgery including MCI-186 / Edaravone (intravenous) treatment, and vehicle group, and 8 rats per group for sham operation group). Rats were kept in the animal facility for one week before all procedures. Animal identification numbers were labeled on the cage tags as well as on the tails. See the experimental design shown in Table 7.

[0252] (Pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride, 2-(2-acetoxybenzoyl)oxybenzoic acid (acetylsalsalate), edaravone, and vehicle were administered 1 hour after MCAO surgery (day 0) and on days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14.

[0253] Table 7. Experimental design JPEG2025504447000025.jpg65161

[0254] Prior to MCAO, animals were fasted overnight but allowed free access to water. Under anesthesia, a midline incision was made in the neck to expose the right common carotid artery (CCA), internal carotid artery (ICA), and external carotid artery (ECA). A commercially available monofilament (with silicone coating) was used as an occlusion plug and inserted through the CCA. The occlusion plug was advanced into the CCA 18±0.5 mm beyond the carotid bifurcation. Light resistance indicated that the occlusion plug was adequately lodged within the anterior cerebral artery, thus blocking blood flow to the middle cerebral artery (MCA). After 1 hour, reperfusion was allowed by fully withdrawing the monofilament. Body temperature was maintained at approximately 36.5°C during the surgical procedure using a heating pad. At 2 hours after occlusion, clinical signs were examined by an observer blinded to the treatment groups to confirm neurological deficits. Neurological deficits (see Table 3) were assessed daily thereafter until the 14th day. 14 days after MCAO, the animals were euthanized and the brains were cut into 5 coronal sections (2 mm thick by using a rat brain matrix). Fresh brain sections were stained with 2% triphenyltetrazolium chloride (TTC) solution at 37°C, and then fixed with 4% paraformaldehyde. Photographs of all sections were taken with a digital camera. These digital photographs were transferred to a computer. The infarct volume per brain was obtained after blindly measuring the infarct area (%) for each section using Image-Pro Plus software. Infarct area (%) = (contralateral hemisphere area - ipsilateral non-infarct area) / contralateral hemisphere area, where the relevant swelling and atrophy were corrected by this formula.

[0255] For the analysis of infarct size, One-Way ANOVA followed by Dunnett's multiple comparison test was used to compare the differences between the treatment and vehicle groups. A t-test was used to compare the differences between the positive control and vehicle groups. P<0.05 was considered to indicate a significant difference between groups. Data were expressed as mean±SEM. For the analysis of body weight, a t-test was used to compare the differences between groups at various days after surgery.

[0256] All animals were in good general condition before the study. During the experiment, mortality was 5 of 25 (20%) in the vehicle group, 5 of 25 (20%) in the edaravone treatment group, 3 of 25 (12%) in the oral acetylsalsalate group, 2 of 25 (8%) in the low dose transdermal (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride group, 1 of 25 (4%) in the medium dose transdermal (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride group, 0 of 25 in the high dose transdermal (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride group, and 0 of 8 (0%) in the sham operation group. Most of the animal deaths occurred between 48 hours (6 rats) and 72 hours after the MCAO operation. All other MCAO animals lost weight between days 1 and 4 after surgery and then maintained a relatively stable health status until euthanasia. Animals in the sham-operated group lost weight only during the first 2 days after sham surgery and then gradually returned to normal levels.

[0257] Several animals with obvious bleeding during surgery were abandoned. After the final stage of the surgery (removal of the occluding plug), the animals were closely monitored until they had fully regained consciousness.

[0258] Rats treated with transdermal (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride showed smaller infarct volumes compared to the intravenous edaravone (41.33% ± 2.792), oral acetylsalsalate (39.25% ± 2.981), and vehicle groups (43.98% ± 2.256), expressed as mean ± SEM: low dose group (36.87% ± 2.265, P < 0.10), medium dose group (35.39% ± 2.479, P < 0.07), and high dose group (32.98% ± 2.546, P < 0.05). The difference of the high dose group against the vehicle group was statistically significant (P < 0.05). In contrast, acetylsalsalate-treated and MCI-186 / edaravone-treated rats showed little reduction in infarct volume. Rats in the Sham group showed unavoidable measurement variation (3.56%±1.257). Most of the infarcts in the vehicle-, acetylsalsalate-, and MCI-186 / Edaravone-treated groups were typically involved in the striatum and frontoparietal cortex supplied by the middle cerebral artery, whereas the infarct size was smaller and the striatum and corresponding cortex were less involved in the low-, medium-, and high-dose groups. Detailed analysis of infarct volume is shown in FIG. 13.

[0259] The body weight of MACO rats dropped dramatically during the first 4 days after surgery and then remained relatively stable. Meanwhile, the body weights of the low (P<0.10), medium (P<0.08), and high (P<0.05) dose groups of (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride increased from day 5 to day 14 and were significantly different from the vehicle group by t-test. Detailed information on the body weight assessment is shown in Figure 14.

[0260] Ischemic injury caused clinical signs of motor dysfunction on the left side. The neurological deficit scores of the high dose (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride group compared with the vehicle group show significant differences. Detailed information on the assessment of neurological deficit scores is shown in Figure 15.

[0261] This study investigated the therapeutic efficacy of (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride against cerebral ischemia in a rat MCAO model. Compared with the vehicle group, the high-dose group of (pyrrolidin-2-yl)methyl 2-(2-acetoxybenzoyl)oxybenzoate hydrochloride showed good neuroprotective effects against cerebral ischemic injury.

[0262] Example 19 Effect of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride on neurological deficits and cerebral infarction in a monkey model of middle cerebral artery (MCA) thrombosis This study was conducted to investigate whether 2-(diethylamino)ethyl acetoxybenzoate hydrochloride improves neurological deficits and cerebral infarction in a monkey model of middle cerebral artery (MCA) thrombosis. The studies in Examples 14-21 are significantly different from other published studies. In the studies in Examples 14-21, drug treatment was initiated 1 hour to 60 days after thrombosis (i.e., formation of a blood clot in a blood vessel), stroke, and heart attack. That is, the treatment is aimed at restoring or improving function after stroke, heart attack, heart failure, or other cardiovascular disease. In contrast, in other published studies, drug treatment was initiated before thrombosis. That is, the drug treatment is aimed at prevention, not recovery or amelioration from the damage of thrombosis.

[0263] Formulations were prepared weekly, stored at 2°C-8°C in amber bottles when not in use, and used within 7 days. Concentrations were expressed as the free base of the test article. A correction factor of 1.13 and purity were used to calculate the free base.

[0264] 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride (15.82 g) was weighed out and added to a 200 ml flask. Approximately 80% of the final volume of 15% (v / v) ethanol in sterile water for injection (vehicle) was added to the vessel and the resulting mixture was stirred until a clear solution was obtained, after which an additional 15% (v / v) ethanol was added to the flask to a final volume of 200 ml (70 mg / ml of 2-(diethylamino)ethyl acetoxybenzoate free base).

[0265] Three hours after the start of the Rose Bengal injection, administration of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, aspirin, or vehicle was started. From the next day through to the 27th day, 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, aspirin, or vehicle was administered twice daily. 2-(diethylamino)ethyl acetoxybenzoate hydrochloride and vehicle were administered transdermally on the back using a disposable syringe. Aspirin was administered orally by gavage using a nasal catheter with a disposable syringe. The catheter was inserted into the stomach via the mouth. Treatment was once on the day of MCA thrombosis (day 0) and twice daily for 27 days. (See experimental design shown in Table 8).

[0266] The monkeys were fed approximately 50-100 g of food per monkey twice daily, along with washed and disinfected fruits or vegetables. Tap water was available ad libitum throughout the study. Fluorescent lighting was provided for approximately 12 hours per day.

[0267] They were fed a certified monkey diet (Beijing Keaoxieli Feed Co., Ltd). The nutritional components of each batch of diet were analyzed. The diet should meet the feed standards according to the national standards of the People's Republic of China GB14924.3-2010 and GB14924.2-2001. Copies of the analysis results were kept in the study file.

[0268] Certified tap water was used. The appearance and microorganisms of the drinking water were analyzed monthly, and toxicological indices (e.g., lead and mercury) were detected annually. The water should meet the drinking water standards according to the National Standard GB5749-2006 of the People's Republic of China. Copies of the analysis results were kept in the study file.

[0269] Table 8. Composition of experimental groups JPEG2025504447000026.jpg40161

[0270] Experimental groups were assigned to 24 monkeys according to a minimization method based on their preoperative body weights.

[0271] Information regarding allocation was disclosed only to those responsible for administration, and was not disclosed to those responsible for creating the MCA thrombosis model, assessing neurological deficits, or measuring infarct lesions. The MCA thrombosis model was performed in a blinded manner, and the operator was not responsible for group allocation and administration.

[0272] Monkeys that had been fasted for at least 12 hours were anesthetized with intramuscular administration of ketamine hydrochloride (20 mg / kg) and placed on a surgical bed. Anesthesia was continued for up to 30 minutes with 1.0% isoflurane in O2 gas. Inhalation anesthesia was administered using an anesthesia machine (SN23402, Hallowell engineering and manufacturing corporation, USA) and a ventilator (SN23402, Hallowell engineering and manufacturing corporation, USA), and the monkeys were placed on a surgical bed. During anesthesia, the body temperature (rectal temperature) of the animals was controlled between 37.0 ± 0.5 °C by a heating mattress (XMTA-7000, Delixi Electric., Ltd.).

[0273] Under anesthesia, the left eye was enucleated using a bipolar coagulation device (Valleylab Force 1C, USA). A craniotomy was then performed at the superolateral side of the left optic nerve. The next day, the animals were anesthetized with ketamine hydrochloride (20 mg / kg), and the dura and subarachnoid membrane were opened. MCA thrombosis was induced by photochemical reaction. The proximal part of the MCA trunk was illuminated with green light of wavelength 532 nm by using a xenon lamp (GL532T3-100FC, Shanghai Laser & Optics Century Co., Ltd.). The illumination was directed by a 3 mm diameter optical fiber attached to a micromanipulator. Irradiation was started, and Rose Bengal 20 mg / kg was injected intravenously. Light illumination was continued for 30 min. The dura was then covered with a moist gelatin sponge.

[0274] Penicillin G potassium (100,000 U per animal) was injected intramuscularly daily for 3 days after surgery to prevent postoperative infections. Neurological deficits (see Table 3) were blindly assessed 1, 3, 5, 7, 14, and 28 days after the onset of MCA thrombosis (day 0). Neurological deficits were scored for consciousness, sensory system, motor system, and skeletal muscle coordination, and a total neurological deficit score was calculated by summing the scores for consciousness, sensory system, motor system, and skeletal muscle coordination. Neurological deficit scores were measured by adapting the scoring method described in Kito G, et al. J Neurosci Meth, 2001, 105:45-53.

[0275] The surviving monkeys were deeply anesthetized with sodium pentobarbital (35 mg / kg, intravenous) 28 days after the onset of MCA thrombosis. After cutting the atria of the heart, the brain was perfused with 200 mL of heparinized saline (10 U / mL) via the common carotid artery, followed by 200 mL of 10% formalin neutral buffer. The whole brain was removed and then placed in a pathology specimen preservation pack containing 10% formalin neutral buffer. For each brain sample, coronal sections (thickness: 7 μm to 8 μm) were prepared at 4 mm intervals at 10 sites. The slices at each site were stained with H&E reagent.

[0276] Measurements of cerebral infarction were performed blindly. The infarct on the ischemic side was defined as the necrotic area compared with the contralateral infarct. The infarct area (A) on the cross section of the left cerebral hemisphere was measured by tracing using a computer image analysis system (Image J).

[0277] If the infarct sizes of the two consecutive slices are D and D', the infarct volume using the slice thickness (4 mm) is calculated by the following formula: Infarct volume = {(D+D') × 4} / 2 It is measured in.

[0278] Total infarct volume is calculated by summing the infarct volumes of 10 consecutive sections.

[0279] Evaluation items: 1) Neurological Deficit Score (consciousness, sensory system, motor system, and skeletal muscle coordination) and Total Neurological Deficit Score 2) Infarct volume

[0280] result: Mortality rates in the vehicle, aspirin, low-dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, and high-dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride groups were 4 out of 7, 1 out of 5, 0 out of 5, and 0 out of 7, respectively. Thus, the postmortem neurological deficits of the animals were expressed as a score at the time of death. Severe neurological deficits were observed in the vehicle group 3 days after MCA thrombosis. These neurological deficits did not show spontaneous recovery. Neurological deficits in the aspirin group tended to show slight recovery up to 28 days after MCA thrombosis. Neurological deficits improved in a dose-dependent manner in the low-dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride and high-dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride groups (see FIG. 16).

[0281] The infarct volumes measured 28 days after MCA thrombosis were 2,873 mm in the vehicle, aspirin, low-dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, and high-dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride groups, respectively. 3 , 1901mm 3 , 945mm 3 , and 988 mm 3 (See FIG. 17.) In the low dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride group and the high dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride group, the infarct volume was significantly reduced.

[0282] In the infarcted region, severe liquefaction was observed in the vehicle group, moderate in the aspirin group and the low-dose 2-(diethylamino)ethylacetoxybenzoate hydrochloride group, and slight in the high-dose 2-(diethylamino)ethylacetoxybenzoate hydrochloride group. In the penumbra region, many mastocyte astrocytes were observed in the high-dose 2-(diethylamino)ethylacetoxybenzoate hydrochloride group, but only a few mastocyte astrocytes were observed in the vehicle group. The aspirin group showed hemorrhage in the penumbra region. In the remote region, delayed neuronal death was observed in all groups, but slight in the 2-(diethylamino)ethylacetoxybenzoate hydrochloride group.

[0283] 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride reduced mortality and infarct volume, and improved neurological deficits. Histopathological findings showed that many mast cell astrocytes were observed in the high-dose group of 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride. Many mast cell astrocytes remained in 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride because of only minor delay in liquefaction and neuronal cell death. In conclusion, 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride was demonstrated to improve ischemic brain damage and delay neuronal cell death in a monkey model of middle cerebral artery thrombosis.

[0284] Example 20 Efficacy study of the effect of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride on myocardial infarction in minipigs Preparation of formulations 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride was dissolved in 15% ethanol to the nominal concentration for transdermal administration (7.91%), and aspirin was dissolved in 0.5% MC to the nominal concentration for oral administration (0.5%).

[0285] 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride and vehicle were administered transdermally to the back using a disposable syringe twice daily from the following day through the 13th day, and aspirin was administered orally by gavage using a nasal catheter with a disposable syringe attached. The catheter was inserted via the mouth into the stomach twice daily from the following day through the 13th day.

[0286] Based on the level of pharmacological effect, dose levels were selected as follows: 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride: Low dose: 15 mg / 0.215 mL / kg (equivalent to 10 mg / kg of aspirin per dose of the free base) twice daily; High dose: 30 mg / 0.430 mL / kg (equivalent to 20 mg / kg of aspirin per dose of the free base) twice daily; Aspirin: 20 mg / 4 mL / kg twice daily.

[0287] Dosage formulations were prepared weekly, stored at 2°C-8°C in amber bottles when not in use, and used within 7 days. Concentrations were expressed as the free base of the test article. A correction factor of 1.13 and purity were used to calculate the free base.

[0288] 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride (15.82 g) was weighed out and added to a 200 ml flask. Approximately 80% of the final volume of 15% (v / v) ethanol in sterile water for injection (vehicle) was added to the vessel and the resulting mixture was stirred until a clear solution was obtained, after which an additional 15% (v / v) ethanol was added to the flask to a final volume of 200 ml (70 mg / ml of 2-(diethylamino)ethyl acetoxybenzoate free base).

[0289] A total of 24 male minipigs were used in this study. At the start of the study, the minipigs were approximately 2-3 weeks old. The minipigs were already quarantined and housed. Before the animals were transferred from the breeding colony, a physical examination was performed and the animals' tolerance to the study was evaluated by a veterinarian. See the experimental design shown in Table 9.

[0290] Table 9. Composition of experimental groups JPEG2025504447000027.jpg39160

[0291] Experimental groups were assigned according to a minimization method based on the preoperative weights of 24 minipigs. The myocardial infarction model was created in a blinded manner, and the operator was not responsible for group allocation and administration.

[0292] The miniature pigs, which had been fasted for at least 12 hours, were anesthetized with intramuscular administration of pentobarbital (25 mg / kg-50 mg / kg) and laid on a surgical bed. Anesthesia was continued with 1.0% isoflurane in O2 gas. Inhalation anesthesia was administered using an anesthesia machine (SN23402, Hallowell engineering and manufacturing corporation, USA) and a ventilator (SN23402, Hallowell engineering and manufacturing corporation, USA), and the animals were laid on a surgical bed. During anesthesia, the body temperature (rectal temperature) of the animals was controlled between 37.0 ± 0.5 °C by a heating mattress (XMTA-7000, Delixi Electric., Ltd.).

[0293] Under anesthesia, a thoracotomy was performed to expose the circumflex coronary artery of the heart. Circumflex coronary artery thrombosis was induced by photochemical reaction. The artery was illuminated with green light of wavelength 532 nm by using a xenon lamp (GL532T3-100FC, Shanghai Laser & Optics Century Co., Ltd.). Illumination was directed by a 3 mm diameter optical fiber attached to a micromanipulator. Illumination was started and 20 mg / kg of Rose Bengal was injected intravenously. Light illumination was continued for 30 min.

[0294] Penicillin G potassium (100,000 U per animal) was injected intramuscularly daily for 3 days after surgery to prevent postoperative infections. Before the myocardial infarction model was created, a telemetry transmitter (TL11M2-D70-PCT, Data Sciences International, Minnesota, USA) was placed inside the jacket, and ECG leads were placed subcutaneously in the right lateral thorax and left lateral abdomen. Physiological signals (ECG) were recorded within 24 hours after administration of Rose Bengal.

[0295] The surviving miniature pig was deeply anesthetized with sodium pentobarbital (30 mg / kg, intravenous) 14 days after the onset of circumflex coronary artery thrombosis. After cutting the femoral artery, a thoracotomy was performed and the heart was removed. The heart was cut into five blocks with a width of 5 mm, and then these blocks were immersed in TTC reagent at 37 °C for 5 min. After staining with TTC reagent, photographs of the five blocks of the heart were taken. All blocks were immersed in 10% formalin neutral buffer. A large necrotic area was seen in the second block, which was cut into sections (thickness: 7 μm to 8 μm). The slices were stained with H&E reagent.

[0296] Measurements of myocardial infarction lesions were performed in a blinded manner. The infarct lesion was defined as the necrotic area of ​​the TTC-stained block. If the infarct sizes of the two consecutive sections were D and D', the infarct volume was calculated using the thickness of the block (5 mm) according to the following formula: Infarct volume = {(D+D') × 5} / 2 was measured.

[0297] Total infarct volume was calculated by summing the infarct volumes of five consecutive sections. Histopathological changes of myocardial infarction (necrosis, inflammation, granulation, etc.) were examined using H&E stained slides.

[0298] Evaluation items 1) Infarct volume 2) Histopathological changes

[0299] result: The infarct volumes measured 28 days after MCA thrombosis were 379 mm in the vehicle, aspirin, low-dose (moderate), and high-dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride groups, respectively. 3 , 431mm 3 , 177mm 3 , and 138mm 3 The low dose group of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride showed moderate and the high dose group of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride showed reduced infarct volume (see FIG. 18).

[0300] Moderate myocardial degeneration and necrosis were observed in the vehicle and aspirin groups, but these histopathological changes were minor in the low-dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride group (moderate) and the high-dose 2-(diethylamino)ethyl acetoxybenzoate hydrochloride group.

[0301] The aim of this study was to evaluate the effect of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride on myocardial infarction in minipigs. ECG assessment showed ST upsegment and abnormal Q waves in all groups, indicating the development of myocardial ischemia after irradiation. On the 28th day after MCA thrombosis, myocardial infarction volume was reduced in the 2-(diethylamino)ethyl acetoxybenzoate hydrochloride group. Thus, this experiment demonstrated that myocardial infarction was ameliorated by repeated administration of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride.

[0302] Example 20. Non-clinical pharmacology and toxicology A series of pharmacology and toxicology studies have been conducted in support of early clinical trials, including a GLP acute dermal maximum tolerated dose study in rats, a GLP acute dermal maximum tolerated dose study in minipigs, a 28-day GLP repeat dose dermal toxicity and toxicokinetic study with 14-day recovery in rats, a 28-day GLP repeat dose dermal toxicity and toxicokinetic study with 14-day recovery in minipigs, a GLP bacterial reverse mutation assay (Ames), a GLP in vitro chromosomal aberration assay in CHO-WBL cells, a GLP in vivo bone marrow micronucleus assay in rats, behavioral effects in rats using a functional observational global assessment, a respiratory safety pharmacology study in rats, a cardiovascular telemetry study in unrestrained conscious minipigs, a dermal irritation study in rabbits, and a sensitization study in guinea pigs.

[0303] Results of the study indicate that 2-(diethylamino)ethyl acetoxybenzoate hydrochloride is safe and generally well tolerated.

[0304] Example 21 Phase 1 Clinical Study A randomized, double-blind, placebo-controlled, dose-escalation study was conducted to evaluate the safety, tolerability, and PK of single and multiple escalating doses of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride administered as a topical spray application.

[0305] In each of the four dose escalation cohorts, eight subjects were randomized to receive the active treatment and two subjects were randomized to a matching placebo. The dose levels studied are shown in Table 10.

[0306] Table 10. Total dose and number of sprays per dose by cohort JPEG2025504447000028.jpg40125

[0307] Each cohort began with administration of a single dose of investigational drug on day 1 of the cohort's study schedule. Serial blood samples for plasma PK analysis of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride and three metabolites of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, specifically 2-(diethylamino)ethyl 2-hydroxybenzoate hydrochloride, aspirin (acetylsalicylic acid, ASA), and salicylic acid (SA), were collected over a 120-hour post-dose period.

[0308] After a 5-day washout, multiple doses of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride were administered twice daily for 7 days (days 6-12, with only the morning dose administered on day 12). After the final dose of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride administered in the morning of day 12, blood samples for PK analysis were collected over a 120-hour post-dose period.

[0309] Safety data were reviewed prior to initiating multiple dosing within a cohort and prior to dose escalation to a higher single dose. Subsequent higher dose cohorts were initiated in the absence of dose-limiting adverse events (AEs) and laboratory toxicities. Decisions for dose escalation and multiple dosing were based on safety and tolerability assessments and were agreed upon by Techfields Inc., the medical monitor, and the principal investigator (PI).

[0310] The maximum individual dose of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride was 700 mg twice daily (cohort 4), with a maximum daily dose of 1400 mg.

[0311] For single-dose PK analysis, serial blood samples were collected at the following time points after dosing on Day 1: 0 hours (pre-dose), 0.25 hours, 0.5 hours, 0.75 hours, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 4 hours, 5 hours, 6 hours, 8 hours, 10 hours, 12 hours, 18 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, and 120 hours. For multiple-dose PK analysis, serial blood samples were collected after the last morning dose on Day 12 according to the same schedule used for single-dose PK analysis. In addition, daily trough PK samples were collected before the morning dose on Days 7 through 11.

[0312] Safety assessments included monitoring of AEs, vital signs (blood pressure (BP), pulse rate, respiratory rate, and oral temperature), clinical laboratory findings, 12-lead electrocardiogram (ECG), skin irritation assessment, and physical examination (PE) findings. A physical examination and resting 12-lead ECG were performed at screening and on Day 13. Vital signs were assessed at screening, upon admission to the study site on Day -1 and on Day 5, as well as prior to and 1 hour after administration of each dose.

[0313] Laboratory tests (chemistry, hematology, and urinalysis) were performed at screening, on admission on day -1, and on days 4, 13, and 17. Stool guaiac tests were performed at screening and at the end-of-study visit on day 17. Skin irritation assessments were performed prior to administration of each dose and 30 minutes after administration of each dose.

[0314] Conclusion: Safety Conclusion: 1. This study demonstrated that 2-(diethylamino)ethyl acetoxybenzoate hydrochloride was safe and well tolerated in all dose regimens studied: single doses up to 700 mg and multiple doses up to 700 mg twice daily. No clear relationship was found between dose and incidence of TEAEs. 2. Subjects did not experience any serious adverse events (SAEs) or TEAEs leading to study discontinuation. 3. Other than the indicated TEAEs, there were no overall clinically meaningful or significant changes in clinical safety parameters or physical examination findings.

[0315] PK's conclusion: Following administration of single or multiple doses of 1.70 mg to 700 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, plasma concentrations of TF0039 and ASA were below the limit of quantification. After administration of single or multiple doses of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride ranging from 2.70 mg to 700 mg, the maximum plasma concentrations of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride were near the lower limit of quantifiable levels. After administration of single doses of 3.70 mg to 700 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride, salicylic acid was predominant in plasma, and both the AUCt and AUCτ of salicylic acid increased with the dose of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride. max was increased from a dose of 350 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride to a dose of 700 mg. After multiple doses of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride from 70 mg to 700 mg, the T of salicylic acid was max was significantly different from the values ​​obtained from the single dose treatment. max and AUC τA dose-proportional increase was observed in salicylic acid. The terminal half-life of salicylic acid after either single or multiple doses of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride ranged from 23 to 36 hours. 4. Accumulation of salicylic acid at steady state (R_C max and R_AUCτ) are max The range for AUCτ was 1.5 to 2.6, and the range for AUCτ was 2 to 8. 5. Wide variability was observed in the PK parameters for both 2-(diethylamino)ethyl acetoxybenzoate hydrochloride and salicylic acid. 6. 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride is readily and almost completely (>99%) converted to salicylic acid following topical spraying of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride.

[0316] Example 22 Phase 2 Clinical Study The Phase 2 clinical trial was a multicenter, randomized, double-blind (within-dose), placebo-controlled, parallel-group, dose-ranging study to evaluate the efficacy and safety of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride nebulization versus placebo in improving function in patients with ischemic stroke, as measured by dichotomized mRS score (0-2 vs. >2) at week 16 for all randomized patients. In addition, the study was to evaluate the safety and tolerability of multiple doses of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride nebulization at weeks 16 and 32, the efficacy of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride nebulization compared with placebo for improving function in patients with ischemic stroke as measured by dichotomized mRS score (0-2 vs. >2) at week 32 for all randomized patients, and the effect of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride nebulization compared with placebo for improving function in patients with ischemic stroke as measured by improvement from baseline to weeks 16 and 32 in National Institutes of Health Stroke Scale (NIHSS) score or Barthel Index (BI).

[0317] Methodology / Research Design Each patient enrolled in Group A was administered the study drug in a double-blind fashion for 16 weeks, beginning between 3 and 60 days after the onset of stroke symptoms (Day 1 of the study). Patients and / or their caregivers were instructed on how to correctly apply the study drug, administering either the study drug or placebo at 20 sprays per dose, twice daily (approximately every 12 hours) for a total daily dose of 280 mg of study drug. The 20 sprays of study drug were administered as follows: 4 sprays around the neck, 2 sprays on the left shoulder, 2 sprays on the right shoulder, 6 sprays on the chest, 3 sprays on the left leg, and 3 sprays on the right leg. Each spray was sprayed on a different area of ​​the skin to achieve maximum absorption. Patients were to wait at least 5 minutes after the last spray before dressing to allow the skin to dry completely.

[0318] Patients were asked to return to the study site at weeks 4, 8, 12, and 16 for efficacy and safety assessments. After the week 16 assessment, patients began the open-label period of the study the next day and received active treatment in an open-label manner until the end of study (EOS) at week 32. That is, patients who were receiving active treatment will continue to receive active treatment, and patients who were receiving placebo will switch to active treatment for the remaining 16 weeks of the study. At the week 16 visit, patients will receive only one dose of study drug. Patients will return to the study site at weeks 24 and 32 (EOS) for efficacy and safety assessments. If a patient prematurely discontinues the study, an early discontinuation visit will occur. Patients will have a follow-up visit approximately 14 days after the early discontinuation visit or the week 32 (EOS) visit.

[0319] Test product, dose, formulation, and mode of administration: The test product was a 7.91% solution of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in 15% ethanol (7% solution of 2-(diethylamino)ethyl acetoxybenzoate) or placebo, which was administered transdermally as a spray. The 7.91% transdermal spray solution consisted of 1582 mg of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride (equivalent to 1400 mg of 2-(diethylamino)ethyl acetoxybenzoate free base) in 20 mL of 15% ethanol (volume / volume). The spray bottle deposited 100 ml of spray solution and 7 mg of 2-(diethylamino)ethyl acetoxybenzoate free base on the skin per spray. Subjects were to apply each spray to different skin areas around the neck, chest, legs, and arms based on their randomly assigned dose level.

[0320] Dosage and regimen: Patients were randomized in a 1:1:1 ratio into Group A, Group B, or Group C, and further randomized in a 2:1 ratio within each treatment group to receive either 2-(diethylamino)ethyl acetoxybenzoate hydrochloride 140 mg per dose (20 sprays per dose) or placebo in Group A, 70 mg per dose (10 sprays per dose) or placebo in Group B, or 35 mg per dose (5 sprays per dose) or placebo in Group C.

[0321] Treatments were administered twice daily (approximately every 12 hours) from study day 1 through the week 16 visit. Each spray was sprayed onto a different area of ​​skin to achieve maximum absorption.

[0322] The first dose was administered at the investigational site at the week 16 visit. Active treatment was administered in an open-label fashion starting the day after the week 16 visit through the end of the study (week 32). Patient Group A received 20 sprays of study drug as follows: 4 sprays around the neck, 2 sprays on the left shoulder, 2 sprays on the right shoulder, 6 sprays on the chest, 3 sprays on the left leg, and 3 sprays on the right leg twice daily for a total of 40 sprays (280 mg) per day. Each spray was sprayed onto a different area of ​​the skin to achieve maximum absorption. Patient Group B received the study drug as follows: 2 sprays on the front of the neck, 1 spray on the left side of the neck, 1 spray on the right side of the neck, and 6 sprays on the chest near the neck, twice daily for a total of 20 sprays (140 mg) per day. Each spray was sprayed onto a different area of ​​the skin to achieve maximum absorption. Patient Group C received the study drug as follows: 1 spray on the front of the neck, 1 spray on the left side of the neck, 1 spray on the right side of the neck, and 2 sprays on the chest, twice daily for a total of 10 sprays (70 mg per day). Each spray was sprayed onto a different area of ​​the skin to achieve maximum absorption.

[0323] Efficacy assessment Primary Efficacy Endpoint The primary efficacy endpoint was the mRS score at week 16 dichotomized as "success" (0-2) or failure (>2) at week 16 for all randomized patients.

[0324] Modified Rankin Scale score mRS scores were assessed at screening, day 1 (baseline), weeks 4, 8, 12, 16, 24, and 32, at the early discontinuation visit (if applicable), and at follow-up visits.

[0325] The mRS score is a measure of a patient's functional level and is dichotomized into good outcome (score = 0-2) versus poor outcome (score ≥ 2). mRS scores range from 0 (no symptoms) to 6 (death) as follows: 0 = no symptoms at all 1 = Symptoms present but no obvious impairment; able to carry out all usual daily living and activities 2 = Minor impairment; unable to perform all previous activities but able to care for self without assistance 3 = Moderately impaired, needs some assistance, but can walk unaided 4 = Moderate to severe disability, unable to walk unaided and unable to meet one's physical needs unaided 5 = Severely disabled, bedridden, incontinent, requiring constant nursing care and attention 6=death

[0326] Secondary Endpoints Secondary efficacy endpoints include change from baseline in NIHSS, BI, and GOS-E scores at weeks 16 and 32 for all randomized patients, and change from baseline in mRS score at week 32. NIHSS and BI scores will be assessed at screening, day 1 (baseline), weeks 4, 8, 12, 16, 24, and 32. Another secondary endpoint is improvement in atherosclerosis in the carotid arteries by ultrasound from baseline to weeks 16 and 32.

[0327] Additional secondary efficacy endpoints include change from baseline in blood flow and atherosclerosis in the carotid arteries at weeks 16 and 32. These variables will be measured at day 1 (baseline) and at weeks 16 and 32.

[0328] National Institutes of Health Stroke Scale The NIHSS is a longitudinal measure of neurological deficits and is used to objectively grade the severity of ischemic stroke. The scale consists of 11 items summarizing specific abilities such as level of consciousness, LOC questioning, best gaze, visual field, facial paresis, upper limb movement, lower limb movement, limb ataxia, sensation, best speech, dysarthria, extinction phenomenon, and attention disorder (previously neglect), and scores range from 0 to 4, with lower numbers indicating a more normal patient condition.

[0329] Barthel Index The BI is an ordinal scale used to measure a patient's ability to perform 10 individual activities of daily living (ADLs). Each item is scored on a 5-point scale (0, 5, 10, or 15) and the individual items are summed to generate a total score between 0 and 100, where 0 is poor ability and 100 is the best. The lowest score of 0 indicates complete dependence on others for ADLs and the highest score of 100 indicates complete independence in ADLs. Higher scores are associated with a higher likelihood of being able to live at home with some independence. A score of 95 or above is considered excellent.

[0330] Rating: Safety Safety assessments included AEs, vital signs (blood pressure, pulse rate, and oral temperature), clinical laboratories, physical examinations, skin irritation, and ECGs at various time points during the study, as indicated in the study flow chart.

[0331] Adverse events of note include local skin reactions around the treated knee(s), upper gastric pain, GI bleeding, serious cardiovascular side effects (e.g., thrombotic events, myocardial infarction, or stroke), jaundice, elevated liver function tests, and nausea.

[0332] clinical results The clinical results (primary and secondary endpoints) are shown in Tables 11 to 14.

[0333] Table 11. Change in modified Rankin Scale score (mRS) for 2-(diethylamino)ethyl acetoxybenzoate hydrochloride versus placebo JPEG2025504447000029.jpg35162

[0334] The mRS results indicate that 2-(diethylamino)ethyl acetoxybenzoate hydrochloride can improve functional recovery after stroke.

[0335] Table 12. Change in National Institutes of Health Stroke Scale for 2-(Diethylamino)ethyl Acetoxybenzoate Hydrochloride vs. Placebo JPEG2025504447000030.jpg35159NIHSS results show that 2-(diethylamino)ethyl acetoxybenzoate hydrochloride may improve functional recovery after stroke.

[0336] Table 13. Change in Barthel Index for 2-(Diethylamino)ethyl Acetoxybenzoate Hydrochloride vs. Placebo The results of JPEG2025504447000031.jpg36160BI show that 2-(diethylamino)ethyl acetoxybenzoate hydrochloride can improve functional recovery after stroke.

[0337] Table 14. Improvement of atherosclerosis in the cervical arteries by ultrasound examination from baseline to weeks 16 and 32 JPEG2025504447000032.jpg84164

[0338] RCCA (proximal wall): maximum IMT of right common carotid artery (proximal wall), RCCA (distal wall): maximum IMT of right common carotid artery (distal wall), LCCA (proximal wall): maximum IMT of left common carotid artery (proximal wall), LCCA (distal wall): maximum IMT of left common carotid artery (distal wall), RICA (proximal wall): maximum IMT of right internal carotid artery (proximal wall), RICA (distal wall): maximum IMT of right internal carotid artery (distal wall), LICA (proximal wall): maximum IMT of left internal carotid artery (proximal wall), LICA (distal wall): maximum IMT of left internal carotid artery (distal wall).

[0339] The results of ameliorating atherosclerosis indicate that 2-(diethylamino)ethyl acetoxybenzoate hydrochloride can reverse atherosclerosis.

[0340] Safety overview All three doses of 2-(diethylamino)ethyl acetoxybenzoate appeared to be safe and generally well tolerated. As mentioned above, gastrointestinal disturbances are a major problem with all NSAIDs, but in this study there were only 15 very mild morbidities (6 constipation, 3 diarrhea, 1 gastroesophageal reflux, 2 abdominal discomfort, 1 abdominal pain, 1 upper abdominal pain, and 1 nausea), none of which were considered drug-related. Morbidity rates were generally similar across all three treatment groups. No significant upper GI tract ulcer complications (i.e. bleeding episodes, perforation, or gastric outlet obstruction) occurred during the study. Mean and central blood pressures remained unchanged. Even the incidence of skin irritation (a common AE of topical drugs) was very low (6 total morbidities) and mild, due to the simple formulation and the fact that 2-(diethylamino)ethyl acetoxybenzoate is a biologically inactive prodrug when on the external surface of the body.

[0341] Because the 2-(diethylamino)ethyl acetoxybenzoate hydrochloride and highly permeable prodrugs of aspirin and other NSAIDs of the present disclosure are capable of crossing one or more biological barriers, they can be administered locally (e.g., topically or transdermally) to reach locations where current drugs cannot adequately reach, where pathologies occur, such as brain tissue damage from stroke, heart tissue damage from heart attack, heart tissue and blood vessel damage from heart failure, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, aortic aneurysm, peripheral arterial disease, atherosclerosis, and other cardiovascular diseases.

[0342] As shown by reductions in animal model studies and examples of human clinical trials, 2-(diethylamino)ethyl acetoxybenzoate hydrochloride can significantly reduce the signs and symptoms of stroke, myocardial infarction, and / or cardiovascular disease in a dose-responsive manner.

[0343] 2-(Diethylamino)ethyl acetoxybenzoate hydrochloride at 35 mg BID, 70 mg BID, and 140 mg BID all appeared to be safe and generally well tolerated. No drug-related GI disturbances were seen in these studies, although GI disturbances are a major problem with all NSAIDs. No significant upper GI tract ulcer complications (i.e., bleeding episodes, perforation, or gastric outlet obstruction) occurred during these studies. Mean and median blood pressures remained unchanged. Even the incidence of skin irritation (a common AE with topical medications) was very low and mild, due to the simplicity of formulation.

[0344] Although certain embodiments have been described in detail above, they are merely illustrative and not limiting of the present invention. The main features of the present invention can be used in various embodiments without departing from the scope of the present invention. Those skilled in the art will clearly understand that many modifications are possible within the scope of the claims without departing from the teachings of the claims. All such modifications and equivalents are intended to be encompassed within the scope of the claims of the present invention and are covered by the claims. All publications, patents, and other references mentioned herein are incorporated by reference in their entirety for all purposes.

Claims

1. A pharmaceutical composition for the prevention or treatment of cardiovascular disease or a cardiovascular condition, wherein the pharmaceutical composition is administered topically in the prevention or treatment of cardiovascular disease or a cardiovascular condition, and comprises a compound of formula (I): 【Chemistry 1】 (In the formula, R ｘ is selected from H, 2,4-difluorophenyl, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl; R ｙ is selected from H, substituted and unsubstituted alkylcarbonyl, substituted and unsubstituted alkoxycarbonyl, substituted and unsubstituted benzoyl, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl; L １ is O, S, NH, O-CH(L ２ ), O-(CH ２ ) ｎ , O—CH(L ２ )-OC(=O), O-CH(L ２ )-O, S-CH(L ２ )—O, and —O—C(═O)—, where n is an integer selected from 1 to 6; L ２ is independently selected at each occurrence from H, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, and substituted and unsubstituted alkylamino; T is a transport unit containing a protonatable amine group, including a substituted or unsubstituted primary amine group, a substituted or unsubstituted secondary amine group, a substituted or unsubstituted tertiary amine group, or a substituted or unsubstituted heterocyclyl containing a protonatable nitrogen, and T is selected from Structures W-1, W-2, W-3, W-4, W-5, and W-6: 【Chemistry 2】 You can choose from R, at each occurrence, is independently selected from a bond, substituted and unsubstituted alkylene, substituted and unsubstituted cycloalkylene, substituted and unsubstituted heterocyclylene, substituted and unsubstituted alkenylene, substituted and unsubstituted alkynylene, substituted and unsubstituted arylene, and substituted and unsubstituted heteroarylene; ２ and optionally O, S, or NR ３ can be further replaced by, where R ３ is hydrogen, C １ ~C ６ Alkyl, C ３ ~C ６ cycloalkyl, or C ６ ~C １０ is aryl, R １ and R ２ are independently selected from H, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocyclyl, substituted and unsubstituted alkyloxyl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl, or alternatively, R １ and R ２ taken together with the nitrogen atom to which they are attached form an optionally substituted heterocyclyl, which optionally further contains one or two additional heteroatoms independently selected from O, S, and N; R １１ , R １２ , and R １３ each independently represents a bond, an optionally substituted C １ ~C ４ alkylene, or optionally substituted C ２ ~C ４ alkenylene, wherein said alkylene and said alkenylene are optionally O, S, or NR ３ One CH replaced by ２ having a group, Here, the R in Structure W-2, Structure W-3, or Structure W-5 １ Any of the adjacent R １１ can be taken together with the nitrogen atom to which they are attached to form an optionally substituted heterocyclic ring, which can optionally further contain one or two additional heteroatoms independently selected from O, S, and N; Here, the R in Structure W-2, Structure W-4, Structure W-5, or Structure W-6 １１ and the R １２ Or the R １１ and the R １３ are optionally connected by an optionally substituted alkylene bridge, and wherein HA is selected from the group consisting of none and pharmaceutically acceptable acids including hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid, bisulfuric acid, phosphoric acid, phosphorous acid, phosphonic acid, isonicotinic acid, acetic acid, lactic acid, salicylic acid, citric acid, tartaric acid, pantothenic acid, bitartaric acid, ascorbic acid, succinic acid, maleic acid, gentisic acid, fumaric acid, gluconic acid, glucaronic acid, saccharic acid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and pamoic acid. or a pharmaceutically acceptable salt thereof, and a stereoisomer thereof, as well as a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of a cardiovascular disease or condition.

2. The compound is 【Transformation 3】 【Chemistry 4】 2. The pharmaceutical composition for preventing or treating cardiovascular diseases or cardiovascular conditions according to claim 1, wherein the compound is selected from the group consisting of benzodiazepine, ...

3. A pharmaceutical composition for the prevention or treatment of cardiovascular diseases or cardiovascular conditions as described in claim 1, wherein the dosage form of the pharmaceutical composition is selected from transdermal patches, creams, foams, gels, lotions, ointments, pastes, powders, shake lotions, solids, sponges, tapes, tinctures, vapors, drops, rinses, sprays, and liquids.

4. A pharmaceutical composition for the prevention or treatment of cardiovascular disease or cardiovascular pathology described in claim 1, wherein the solution of the pharmaceutical composition is selected from an alcohol solution, an acetone solution, a dimethyl sulfoxide solution, an aqueous alcohol solution, an aqueous acetone solution, and an aqueous dimethyl sulfoxide solution.

5. A pharmaceutical composition for the prevention or treatment of cardiovascular disease or cardiovascular pathology described in claim 1, wherein the solution of the pharmaceutical composition contains about 5% (volume / volume) to about 50% (volume / volume) aqueous ethanol solution.

6. A pharmaceutical composition for the prevention or treatment of cardiovascular disease or cardiovascular pathology described in claim 1, wherein the concentration of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in the pharmaceutical composition is from about 10 mg / mL to about 200 mg / mL.

7. A pharmaceutical composition for the prevention or treatment of cardiovascular disease or cardiovascular pathology described in claim 1, wherein the concentration of 2-(diethylamino)ethyl acetoxybenzoate hydrochloride in the pharmaceutical composition is from about 10 mg / g to about 200 mg / g.

8. A pharmaceutical composition for the prevention or treatment of cardiovascular disease or cardiovascular pathology described in claim 1, wherein the pharmaceutical composition has a unit dose of about 0.01 mL to about 1 mL.

9. 2. The pharmaceutical composition for preventing or treating cardiovascular diseases or cardiovascular conditions according to claim 1, wherein the cardiovascular disease or cardiovascular condition is one or more selected from the group consisting of stroke, angina pectoris, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, peripheral arterial disease, atherosclerosis, and other cardiovascular diseases.

10. 2. The pharmaceutical composition for preventing or treating cardiovascular disease or cardiovascular conditions described in claim 1, wherein the topical administration comprises applying the pharmaceutical composition to the skin surface of the subject in at least one area selected from the neck, chest, back, abdomen, head, arms, hands, legs, feet, and combinations thereof.

11. 2. The pharmaceutical composition for preventing or treating a cardiovascular disease or condition according to claim 1, wherein the local administration comprises applying the pharmaceutical composition to the subject in a single dose per site, and the one or more unit doses are from 1 unit dose to 200 unit doses.

12. 2. The pharmaceutical composition for preventing or treating a cardiovascular disease or condition according to claim 1, wherein the topical administration comprises applying the pharmaceutical composition to the subject in a dosage regimen selected from once a day, twice a day, three times a day, four times a day, five times a day, six times a day, seven times a day, and eight times a day.

13. 2. The pharmaceutical composition for preventing or treating a cardiovascular disease or condition according to claim 1, wherein the topical administration comprises applying the pharmaceutical composition to the subject at a dosage regimen selected from once every hour, once every 2 hours, once every 3 hours, once every 4 hours, once every 5 hours, once every 6 hours, once every 7 hours, once every 8 hours, once every 9 hours, once every 10 hours, once every 11 hours, once every 12 hours, once every 13 hours, once every 14 hours, once every 15 hours, once every 16 hours, once every 17 hours, once every 18 hours, once every 19 hours, once every 20 hours, once every 21 hours, once every 22 hours, once every 23 hours, and once every 24 hours.

14. The pharmaceutical composition for preventing or treating cardiovascular diseases or cardiovascular conditions according to claim 1 , wherein the subject is a human.

15. 15. The pharmaceutical composition for preventing or treating a cardiovascular disease or condition according to any one of claims 1 to 14, wherein said topical administration comprises applying said pharmaceutical composition to said subject for a period ranging from one day to a lifetime.

16. 15. A kit for preventing or treating a cardiovascular disease or condition, comprising the pharmaceutical composition of any one of claims 1 to 14, wherein the pharmaceutical composition is adapted for topical administration to one or more sites of the subject in an amount of from about 1 mg to about 7200 mg per day.

17. 17. The kit of claim 16, wherein the pharmaceutical composition is configured so as to be topically administered to the subject in an amount of about 1 mg to about 700 mg per dose, once a day, twice a day, three times a day, or four times a day.

18. The kit according to claim 16, wherein the pharmaceutical composition is provided in a set that can be topically administered to the subject in an amount of about 1 mg to about 200 mg per spray per site.

19. The pharmaceutical composition is applied to 1 cm of skin per site. ２ Approximately 5 μg per 1 cm of skin ２ 17. The kit of claim 16, wherein the kit is configured to be topically administered to the subject in an amount of about 7 mg per unit area.

20. 17. The kit of claim 16, wherein the cardiovascular disease or cardiovascular condition is at least one selected from the group consisting of stroke, angina pectoris, myocardial infarction, heart failure, coronary artery disease, rheumatic heart disease, hypertensive heart disease, atrial fibrillation, congenital heart disease, endocarditis, peripheral arterial disease, atherosclerosis, and other cardiovascular diseases.

21. 17. The kit of claim 16, wherein the subject is a human.

22. 14. A method for preventing or treating a cardiovascular disease or condition, comprising topically administering to a subject the pharmaceutical composition of any one of claims 1 to 13, wherein the subject is not a human.

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