Compositions and methods for enhancing efficacy of dicarboxylic esters and uses of such compositions

Abstract

The present disclosure provides methods and compositions effective to enhance the therapeutic efficacy of a dicarboxylic acid ester, the compositions comprising a dicarboxylic acid ester and at least one solvent. In some embodiments, the composition further comprises at least one terpene, and other embodiments include methods of preparing the composition and the use of the composition for treating various diseases treatable with dicarboxylic acid esters.

Description

Technical Field

[0001] This disclosure relates to methods and pharmaceutical compositions that can effectively enhance the therapeutic effects of dicarboxylic acid esters, the pharmaceutical compositions comprising a dicarboxylic acid ester and at least one solvent. In some embodiments, the compositions further comprise at least one terpene, and other embodiments include methods for preparing the compositions and the use of the compositions for treating a variety of diseases treatable with dicarboxylic acid esters. Background Technology

[0002] Diethyl azelaate (DEA) has shown unexpected versatility in both oral and topical administration, and can be used to treat insulin resistance, musculoskeletal pain, and poisoning caused by phospholipases present in animal venom. See, for example, U.S. Patents Nos. 10,251,857B2, 11,026,912B2, 11,918,555, and 11,911,358, the disclosures of which are incorporated herein by reference in their entirety. See also Streeter RT and Izbicka E, “Diethylazelate for the treatment of brown recluse spider bite, a neglected orphanindication,” In Vivo 36(1):86-93, (2022).

[0003] Anesthetics and analgesics are widely used to treat pain. Recently, Pavel et al. detailed the biochemical mechanisms by which general anesthetics exert their pharmacological effects. Pavel showed that inhaled anesthetics disrupt the order of lipid rafts and induce anesthesia through this disruption. The most well-studied lipid rafts are membrane domains rich in cholesterol and sphingomyelin (e.g., monosialotetrahexosylganglioside 1, akaGM1), which bind to the cholera toxin B subunit (CTxB). The GM1 lipid raft contains phospholipase D2 (PLD2). Inhaled anesthetics increase the mobility of the GM1 lipid raft, thereby disrupting it and causing PLD2 to leave the raft and translocate to the vicinity of TREK1. Once there, PLD2 hydrolyzes phosphatidylcholine (PC) to produce phosphatidic acid (PA) and choline. PA then binds to and activates TREK1. When TREK1 is activated, the resulting potassium influx leads to loss of consciousness and analgesia. Pavel et al. also showed that PLD2 translocation also activates a second type of pathway called TRAAK, which is an anesthesia-insensitive homologue of TREK-1. [Pavel MA, et al., PNAS, (2020) 117(24): 13757-66].

[0004] The inventors have discovered that the dicarboxylic acid esters of this disclosure, when used in combination with at least one solvent and at least one terpene, exhibit surprising effects in treating a variety of ailments, including pain, diabetes, and exposure to brown hermit spider venom and bee venom. The inventors have also discovered that the dicarboxylic acid esters produce a beneficial synergistic effect when used in combination with at least one solvent and at least one terpene.

[0005] Dimethyl sulfoxide (DMSO) is an organosulfur compound that enhances the permeability of plasma membranes containing drugs, DNA, and other substances. Increased DMSO concentrations have been reported to lead to thinning of the plasma membrane, subsequently creating pores, and ultimately causing the bilayer to collapse. [deMenorval MA, et al., “Effects of dimethyl sulfoxide in cholesterol-containing lipid membranes: A comparative study of experiments in silico and with cells,” PLoS One Vol 7(7):e41733,(2012)]. DMSO can provide rapid, moderate, and temporary relief from arthritis pain [Swanson B, "Medical use of dimethyl sulfoxide (DMSO)," Rev Clin Basic Pharm, Vol. 5, pp. 1-33, (1985)], and aids in wound healing; Capriotti K, et al., "Dimethyl sulfoxide: History, chemistry, and clinical utility in dermatology," J Clin Aesthet Dermatol, Vol. 5(9): 24-26, (2012)]. DMSO applied to the skin quickly produces a distinctive garlic flavor on the tongue, indicating its rapid penetration through tissues and distribution throughout the body. [Horita A, et al., "Skinpenetrating property of drugs dissolved in dimethyl sulfoxide (DMSO) and other vehicles," Live Sciences, Vol. 3, pp. 1389-1395, (1964)]. The oral use of DMSO is limited to the treatment of systemic amyloid A amyloidosis, a complication of a chronic inflammatory disease.

[0006] Terpenes can also enhance skin permeability by interacting with intercellular lipids in the stratum corneum (the outermost layer of skin). Even at low concentrations, terpenes can enhance the activity of both hydrophilic and lipophilic drugs. [Chen J, et al., “Naturalterpenes as penetration enhancers for transdermal drug delivery,” Molecules, vol. 21, pg. 1709, (2016)]. Representative terpenes, such as limonene and menthol, exhibit low cytotoxicity in vivo and can improve the bioavailability of drugs in animal models and human skin. Limonene has anti-inflammatory activity when applied to the skin and promotes wound healing in vivo. [d'Alessio PA, et al., “Skin repair properties of d-limonene and perillyl alcohol in murine models,” Antiinflamm AntiallergyAgents Med Chem, Vol 13(1):pp. 29-35, (2014)]. Menthol can stimulate skin nociceptors, initiate the release of vasodilatory peptides, and increase skin temperature.

[0007] At the most basic level, all types of pain, except psychogenic pain, involve the transmission of aversive or nociceptive stimuli to the central nervous system via afferent nerves. The transmission of nerve impulses must occur at multiple points in the transmembrane signaling chain [Basbaum A, et al., Cell. (2009) 139(2): 267-84]. The role of the plasma membrane in pain mechanisms has been demonstrated in vivo by increasing lipid raft mobility through cholesterol consumption by cyclodextrins, thereby disrupting nerve signaling and reversibly reducing prostaglandin E2 (PGE2)-induced hyperalgesia [Ferrari, L, et al., J. Pain (2015) 16(1): 60-6].

[0008] Most pain conditions in human and animal models of pain result in significant alterations in mechanosensory sensation, whereby mechanical forces initially act on the plasma membrane. A study on the link between mechanical stimulation and membrane-associated phospholipase D2 (PLD2) signaling provides clues to further elucidate this process. PLD2 is a mechanosensitive enzyme located at a membrane lipid site composed of cholesterol, ganglioside GM1, and the mechanoactivated ion channel TREK-1, which is responsible for downstream signaling. [Petersen EN, et al., “Mechanical activation of TWIK-related potassium channel by nanoscopic movement and rapid second messenger signaling,” Elife, Vol. 12 (2024)]. A commonly used experimental method for measuring the level of stimulation-induced skin sensitivity is the use of a set of graded monofilaments known as von Frey fibers. [Mills C, et al., "Estimating efficacy and drug ed50's using von Freythresholds: Impact of Weber's law and log transformation," J Pain, Vol. 13(6), pp. 519-523, (2012)].

[0009] Other members of the phospholipase family, represented by phospholipase A2 (PLA2), are expressed in a variety of species, from bacteria to humans. PLA2 can induce pain and inflammation, and in some cases, it also exhibits hemolytic activity. Inhibition of PLA2 has been considered a therapeutic target. [Yedgar S, et al., “Inhibition of phospholipase A(2) as a therapeutic target,” Biochim Biophys Acta, Vol. 1488(1-2), pp. 182-187, (2000)]. DEA and its associated azelate have been reported to inhibit the enzymatic and hemolytic activity of PLA2 in bee and snake venom. [Streeper RT and Izbicka E, “Diethyl azelate for the treatment of brown recluse spider bite, aneglected orphan indication,” In Vivo 36(1):86-93, (2022)].

[0010] The inventors have also demonstrated that dicarboxylic esters can induce a variety of changes in cellular and biological signaling in therapeutically useful ways. In addition to their other pharmacological activities, dicarboxylic esters also induce the release and / or production of signaling molecules with analgesic properties. Reactive cellular molecules include leptin, macrophage granulocyte colony-stimulating factor (M-CSF), granulocyte colony-stimulating factor (G-CSF), and granulocyte-macrophage colony-stimulating factor (GM-CSF). In addition, the inventors have found that treatment of mammalian cells and tissues with dicarboxylic acid esters can inhibit the production of pain-related markers, including inducible nitric oxide synthase (iNOS) [Kwok Y, et al., PLZhang J, et al., Int. Anesthesiol. Clin. (2007) 45(2): 27-37], prostaglandin E2 (PGE2) [Ahlawat A, et al., Eur. J. Pharmacol. (2018) 818: 419-28] and adenosine triphosphate (ATP) [Mense S, Dtsch Arztebl Int 2008; 105(12): 214–9].

[0011] There is a need in the art to enhance the efficacy of dicarboxylic acid esters (including diethyl azelaate) in treating pain, lowering blood glucose levels, inhibiting hemolysis and / or related pain caused by PLA2, PLD, and other phospholipases represented by PLA2, and other diseases and conditions. The embodiments described herein are intended to address and meet these needs, as well as other needs that will become apparent to those skilled in the art upon reading the embodiments described below. Summary of the Invention

[0012] This disclosure provides methods and pharmaceutical compositions comprising a substance having formula I: R2OOC-(CH2) n -COOR1 dicarboxylic acid ester, at least one solvent and at least one terpene,

[0013] Where n is between 4 and 10, and each R1 and R2 is independently a lower alkyl group. Some embodiments of this invention involve inducing analgesia in subjects, thereby inhibiting pain that is partly caused by activation of pattern recognition receptors (PRRs) including Toll-like receptors (TLRs), nucleotide oligomer domain (NOD) receptors, and / or Dectin receptors.

[0014] Some embodiments described herein involve inducing analgesic effects by promoting the secretion of leptin, macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and / or granulocyte colony-stimulating factor (G-CSF), thereby suppressing local pain in the subject.

[0015] In some aspects, embodiments of this document relate to methods and pharmaceutical compositions for inducing analgesic effects by inhibiting the expression and secretion of prostaglandin E2 (PGE2) and / or inducible nitric oxide synthase (iNOS) and / or adenosine triphosphate (ATP), thereby inhibiting local pain in a subject, said pharmaceutical composition comprising a dicarboxylic acid ester of formula I, at least one solvent, and at least one terpene.

[0016] In some respects, embodiments of this document relate to a method for inducing analgesia to suppress localized pain in a subject, the method comprising the step of administering to a subject in need a composition comprising a dicarboxylic acid ester of formula I, at least one solvent, and at least one terpene, wherein the administration results in a reduction of symptoms associated with the pain condition.

[0017] In some respects, embodiments of this document relate to methods for treating type 2 diabetes, insulin resistance, and other diseases caused by high blood sugar levels by administering to a subject in need a composition comprising a dicarboxylic acid ester of formula I, at least one solvent, and at least one terpene, wherein the administration results in a reduction of blood sugar levels.

[0018] In some respects, embodiments of this document relate to methods of treating disorders, diseases, and conditions associated with PLA2 and / or PLD having pathological exogenous or endogenous phospholipase activity by administering to a subject in need a composition comprising a dicarboxylic acid ester of formula I, at least one solvent, and at least one terpene, wherein the administration results in the inhibition of PLA2 or PLD-induced hemolysis.

[0019] In some aspects, embodiments of this document relate to the use of a dicarboxylic acid ester of formula I, at least one solvent, and at least one terpene in the preparation of a medicament for inducing analgesia, thereby inhibiting local pain in a subject. In some aspects, embodiments of this document relate to the use of a composition comprising a dicarboxylic acid ester of formula I, at least one solvent, and at least one terpene in lowering blood glucose levels, thereby treating type 2 diabetes, insulin resistance, and other disorders, diseases, or conditions caused by elevated blood glucose levels.

[0020] In some respects, embodiments thereof relate to the use of compositions comprising a dicarboxylic acid ester of formula I, at least one solvent, and at least one terpene in the preparation of medicaments for inhibiting hemolysis (including hemolysis induced by PLA2 and / or PLD in a subject's body), thereby treating diseases and conditions caused or associated with PLD and / or PLA2. Attached Figure Description

[0021] The following figures illustrate certain aspects of this disclosure. A better understanding of this disclosure can be achieved by referring to one or more of these figures in conjunction with the detailed description of the specific embodiments presented herein.

[0022] Figure 1 This study demonstrates the effect of analgesic effect on the duration of sensitivity response when used in combination with DEA ​​in a skin mechanosensitivity test.

[0023] Figure 2 A-2B shows the dose-response of limonene used in combination with DEA ​​and DMSO, assessed by the duration of sensitivity inhibition in a skin mechanosensitivity test. Figure 2 A shows the dose-response of limonene relative to a corrected baseline time. Figure 2 B shows the dose-response of limonene relative to the corrected baseline time AUC.

[0024] Figure 3 A-3B shows the effects of several relevant medium-chain fatty acid diesters, used alone with DMSO (0.5M each) and in combination with 2% limonene, on the duration of sensitivity inhibition in a skin mechanosensitivity test. Figure 3 A shows the effects of several related medium-chain fatty acid diesters on the time to reach baseline when used alone with DMSO (0.5M each) and in combination with 2% limonene; Figure 3 B shows the effect of several related medium-chain fatty acid diesters on the AUC at baseline time when used alone with DMSO (0.5M each) and in combination with 2% limonene.

[0025] Figure 4 The effects of DEA, DMSO, limonene, and menthol as single agents and in combination on the duration of sensitivity inhibition were demonstrated in a skin mechanosensitivity test.

[0026] Figure 5 The superiority of oral administration of the mixture of 77 / 21 / 2DEA / DMSO / limonene over DEA alone in improving blood glucose levels in diabetic subjects was demonstrated.

[0027] Figure 6 The combination of 77 / 21 / 2DEA, DMSO, and limonene demonstrated superiority over the use of a single drug in inhibiting hemolysis induced by phospholipase A2. Detailed Implementation

[0028] All applications, publications, patents and other references cited in this article are incorporated herein by reference in their entirety.

[0029] As used herein, the singular forms “a,” “an,” and “the” include plural indicators unless the context clearly indicates otherwise. The term “or” refers to a single element or a combination of two or more elements among the alternative elements, unless the context clearly indicates otherwise. As used herein, “comprising” means “including.” Therefore, “comprising A or B” means “including A, B, or A and B,” without excluding additional elements.

[0030] A range may be expressed in this document as from “about” a particular value and / or to “about” another particular value. The term “about” as used herein is intended to define the numerical value it modifies, indicating that such a value is a variable within a range of error. When no specific range of error is stated, such as the standard deviation of the mean given in a data chart or table, the term “about” should be understood to mean the range including the stated value as well as the range encompassed by rounding to that number (taking into account significant figures).

[0031] As used herein, all numerical values ​​or ranges include integers within the range as well as values ​​or fractions of integers within the range, unless the context explicitly indicates otherwise. Furthermore, such ranges are intended to include the numbers themselves and any subranges between them. The range can be integers or consecutive between (including) the endpoints. Thus, for example, a range of approximately 3 hours to approximately 10 hours includes 3 hours, 4 hours, 5 hours, etc., and 3 hours 1 minute, 3 hours 2 minutes, 3 hours 4 minutes, etc., 4 hours 1 minute, 4 hours 2 minutes, 4 hours 4 minutes, etc. A range of 90-100% includes 92.2% to 97.5%, 91.5% to 94.5%, etc. A range of 1 to 25 days includes subranges from 1 day to 5 days, or from 3 days to 7 days, or from 5 days to 25 days.

[0032] As used herein, the term "comprising" means that a pharmaceutical composition (or composition) and method includes the listed elements, but does not exclude other elements. When applied to compositions according to embodiments of the invention, the term "substantially composed of" means that the composition may contain additional elements, provided that these additional elements do not substantially alter the composition. When applied to a composition, the term "substantially alters" means an increase or decrease in the therapeutic efficacy of the composition compared to the efficacy of a composition composed of the elements stated herein. In other words, "substantially composed of" when used to define a composition should indicate the exclusion of other components that are of any essential significance to the composition. Therefore, a composition substantially composed of the components defined herein does not exclude methods of separation and purification as well as trace contaminants in pharmaceutically acceptable carriers. "Comprising of" should mean the exclusion of other component elements in greater than trace amounts and substantial method steps for administering the compositions of the invention. Embodiments defined by each of these transitional terms are within the scope of the invention.

[0033] As used herein, the terms “optional” or “optionally” mean that an event or situation described below may or may not occur, and the description includes both the possibility that the event or situation may occur and the possibility that it may not occur.

[0034] As used herein, "subject" refers to mammals, such as primates, and in one implementation, humans. Non-human primates include marmosets, monkeys, chimpanzees, gorillas, orangutans, and gibbons. The term "subject" also includes cats, dogs, ferrets, chinchillas, mice, rabbits, rats, gerbils, guinea pigs, cattle, horses, pigs, sheep, goats, chickens, turkeys, ducks, pheasants, pigeons, doves, parrots, cockatoos, geese, etc.

[0035] As used herein, the “subject in need” of the methods disclosed herein may be a subject suffering from a pain condition.

[0036] As used herein, the term "therapeuticly effective" amount refers to an amount of active ingredient sufficient to induce local analgesia in a subject. Therapeuticly effective amounts of the pharmaceutical compositions of this disclosure can effectively disrupt lipid rafts. Therapeuticly effective amounts of the pharmaceutical compositions of this disclosure can effectively upregulate (e.g., stimulate) the expression and / or secretion of leptin, M-CSF, G-CSF, GM-CSF, etc., in the cells of a subject. Therapeuticly effective amounts of the pharmaceutical compositions of this disclosure can also effectively downregulate (e.g., inhibit) the expression and / or secretion of ATP, iNOS, PGE2, etc., in the cells of a subject. In other words, a "therapeuticly effective" amount refers to an amount that can alleviate, reduce, decrease, or stabilize at least one clinical condition in a subject to a certain degree. Those skilled in the art will understand that the therapeutic effect does not need to be complete or curative, as long as it provides some benefit to the subject. The effective amount can vary depending on factors such as the wound or affected area, the disease or condition being treated, the specific targeted construct being administered, the subject's body size, or the severity of the disease or condition. Those skilled in the art can determine the effective amount of a particular composition empirically without excessive experimentation.

[0037] As used herein, the term "active ingredient" refers to a bioactive substance. In embodiments, the dicarboxylic acid ester of this disclosure is the active ingredient in the pharmaceutical composition. In embodiments, the dicarboxylic acid ester of this disclosure is the sole active ingredient in the pharmaceutical composition. In embodiments, the azelaic acid ester of this disclosure is the active ingredient in the pharmaceutical composition. In embodiments, diethyl azelaic acid is the active ingredient in the pharmaceutical composition.

[0038] As used in this article, the term "pharmaceutically acceptable carrier" means any suitable adjuvant, carrier, excipient, or stabilizer, which may be in solid or liquid form, such as tablets, capsules, powders, solutions, suspensions, or emulsions, that does not cause significant irritation to the subject and does not eliminate the biological activity and properties of the applied active ingredient.

[0039] The term “disease” as used in this article is intended to be generally synonymous with the terms “symptom” and “condition” (as in medical condition) and to be used interchangeably with them, as they all reflect an abnormal condition of the human or animal body, or a part of it that impairs normal function, usually manifested as obvious signs and symptoms, and resulting in a shortened lifespan or reduced quality of life in humans or animals.

[0040] As used herein, “treating” or “treatment” for a disease or condition can refer to preventing the disease or condition, slowing the onset or progression of the disease or condition, reducing the risk of the disease or condition developing, preventing or delaying the development of symptoms associated with the disease or condition, alleviating or ending symptoms associated with the disease or condition, producing complete or partial remission of the disease or condition, or a combination thereof. Treatment can also refer to prophylactic or preventative treatment of the disease or condition.

[0041] As used herein, the term “inhibition” refers to a specific effect, function, interaction, or occurrence of a symptom that reduces, lowers, limits, and / or blocks. The terms “inhibition,” “reduction,” and “lowering” are used interchangeably herein. In embodiments, the term refers to reducing or preventing the level of a specific activity, function, interaction, or occurrence of certain symptoms in a subject (e.g., the level of a biomarker, including but not limited to protein or non-protein molecules in tissues and body fluids), by a reduction of at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or less than the amount in the corresponding control group. For example, pain is reduced, lowered, or inhibited if a pain indicator (e.g., blood levels of prostaglandin PGE2) is reduced by at least about 10%, 20%, 30%, 50%, 80%, or 100%. In the implementation scheme, compared with the pain prior to the administration of the dicarboxylic acid ester of this disclosure, the pain is reduced, decreased, or suppressed by at least about 1 / 2, 2 / 3, 3 / 4, 4 / 5, or more. The measurable change can be objective (e.g., measurable by some test or marker, such as in vitro or in vivo determination, testing, or observation) or subjective (e.g., the subject gives an indication of the effect or feels the effect).

[0042] As used in this article, the term "pain" refers to an unpleasant sensory and emotional experience associated with or described in relation to actual or potential tissue damage, and includes sensations of more or less localized discomfort, distress, or pain resulting from stimulation of specific nerve endings. Pain can be of various types, including but not limited to nociceptive pain, neuropathic pain, or nociplastic pain, whether acute or chronic. The goal of inducing analgesia in subjects is to reduce the degree or severity of pain perceived by the subjects.

[0043] As used herein, the term "localized pain" refers to non-systemic pain experienced in a specific location or area of ​​a particular part of the subject's body. In the implementation plan, the area of ​​pain is associated with the primary lesion.

[0044] As used herein, the term "venom poisoning" means contact with or injection of venom into a subject due to a bite, fang, tooth, or sting from a venom carrier. The term "venom poisoning" also refers to injection from a venom delivery device. As used herein, the term "venom" means any toxic or poisonous substance that is transmitted subcutaneously, intramuscularly, or locally to a subject through bite, sting, or contact with a venomous animal; such substance may contain various toxins, such as, but not limited to, hemotoxins, cytotoxins, myotoxins, and neurotoxins. As used herein, the term "venom carrier" means any animal or organism that produces, secretes, or carries venom. Examples of venomous animals include, but are not limited to, insects, reptiles, amphibians, arthropods, mollusks, cnidarians, coelenterates, or other venomous vertebrates or invertebrates. Examples of venomous organisms include, but are not limited to, bacteria or fungi.

[0045] The term “necrotic arachnid poisoning” as used in this article is generally synonymous with and can be used interchangeably with terms such as “necrotic arachnid poisoning”, “necrosis”, “skin necrosis”, or “tissue necrosis”, referring to local skin and tissue damage caused by venom poisoning or contact with PLD toxin.

[0046] As used in this article, the term "skin ulcer" or "ulcer" refers to an open sore or wound on the skin in which the epidermis is missing or otherwise damaged. The underlying dermis or subcutaneous tissue may be exposed. The surrounding skin may be red and inflamed. The main symptoms and signs of any inflammatory process are pain, redness, heat, swelling, tenderness, and loss of function. Such open ulcers are susceptible to infection by pathogens such as bacteria, fungi, and viruses. In advanced cases, the ulcer may ooze pus. The pus (dead immune cells, skin cells, subcutaneous tissue cells, cytofluic fluid, and infectious agents) accumulates within the cavity of the skin ulcer, forming an abscess.

[0047] Diabetes is a group of metabolic diseases characterized by high blood sugar (glucose) levels caused by defects in insulin secretion or action, or both. Type 2 diabetes, or T2D, refers to one of the two main types of diabetes. In this type, pancreatic beta cells produce insulin at least in the early stages of the disease, but the body cannot effectively use insulin because its cells become resistant to its action. Later in the disease, the beta cells may stop producing insulin altogether. Type 2 diabetes is also known as insulin-resistant diabetes, non-insulin-dependent diabetes, and adult-onset diabetes.

[0048] "Prediabetes" refers to one or more early diabetes-related conditions, including impaired glucose utilization, abnormal or impaired fasting blood glucose levels, impaired glucose tolerance, impaired insulin sensitivity, and insulin resistance.

[0049] Insulin resistance refers to a condition where cells become resistant to the action of insulin (a hormone that regulates glucose uptake by cells), or produce insufficient amounts of insulin to maintain normal blood sugar levels. Cells become less responsive to insulin's role in transporting glucose from the bloodstream to muscles and other tissues (i.e., reduced insulin sensitivity). Eventually, the pancreas produces far more insulin than normal, and cells develop resistance. As long as enough insulin is produced to overcome this resistance, blood sugar levels can remain normal. Once the pancreas is unable to maintain insulin secretion, blood sugar begins to rise, leading to diabetes. Insulin resistance ranges from normal (insulin sensitive) to insulin resistance (IR).

[0050] Obesity is a chronic condition characterized by an excess of body fat. Normal body fat percentage (expressed as a percentage of body weight) is 25-30% for women and 18-23% for men. Obesity is defined as a body fat percentage exceeding 30% for women and 25% for men.

[0051] Extensive literature on tissue penetration enhancers rarely addresses the direct enhancement of therapeutic efficacy; it typically focuses only on increasing drug concentrations in tissues, generally assuming that increased concentration correlates with enhanced efficacy. The inventors unexpectedly discovered that permeability and efficacy are not linearly related. For example, when DEA is used in combination with solvents such as DMSO, the efficacy exhibits a bell-shaped curve, with a sharp peak at a solvent concentration of approximately 21%, and significantly reduced efficacy below and above this concentration. Such a pronounced peak would not have been observed if permeability were the sole driver of enhanced efficacy. While not bound by any operational or functional theories, the inventors propose that for DEA / solvent / terpene combinations, efficacy is enhanced within a narrow and specific range of DEA / solvent molar ratios, and that the efficacy of preferred DEA / solvent ratios is further enhanced by preferred amounts of at least one terpene (e.g., limonene and menthol). The embodiments described herein demonstrate that permeability enhancement does not play a substantial role in enhancing DEA efficacy. Pharmaceutical Composition

[0052] In its embodiments, this disclosure provides methods and pharmaceutical compositions for inducing analgesia to suppress pain in a subject, including administering to the subject a pharmaceutically acceptable composition having formula I: R2OOC-(CH2). n A pharmaceutical composition comprising a dicarboxylic acid ester of COOR1, at least one solvent, and at least one terpene.

[0053] In the embodiments, n is 4 to 10, 6 to 9, or 7 to 8. In the embodiments, each R1 and R2 is independently a lower alkyl group. The term "lower alkyl group" as used herein refers to a C1 to C6 saturated straight-chain or branched alkyl group. Examples of suitable lower alkyl groups (R1 and R2) in Formula I include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, etc. In the embodiments, R1 and R2 are the same. In the embodiments, R1 and R2 are different.

[0054] In this embodiment, the dicarboxylic acid ester is azelaic acid ester. In this embodiment, the azelaic acid ester is diethyl azelaate, diisopropyl azelaate, or a mixture thereof. In this embodiment, the azelaic acid ester is diethyl azelaate.

[0055] In one embodiment, the dosage of the dicarboxylic acid ester of Formula I is sufficient to induce analgesia in a subject, thereby reducing the pain response. In another embodiment, the dosage of the dicarboxylic acid ester of Formula I may be sufficient to reduce the pain response, for example, by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%. In yet another embodiment, the dosage of the dicarboxylic acid ester of Formula I is sufficient to lower the blood glucose level in a subject to treat one or more conditions, diseases, or illnesses (e.g., insulin resistance, type II diabetes) caused by elevated blood glucose levels. In yet another embodiment, the dosage of the dicarboxylic acid ester of Formula I is sufficient to inhibit hemolysis caused by the toxin, thereby treating conditions, diseases, and illnesses associated with PLD toxin.

[0056] In the embodiments, the dosage of the dicarboxylic acid ester of Formula I is sufficient to provide the above-mentioned effects, and it can be administered in the range of, for example, about 10% to about 90%, about 20% to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 70% to about 80%, or any value or range between therewith.

[0057] The solvent used with the active ingredient dicarboxylic acid ester (preferably diethyl azelate) can be any solvent that enhances the therapeutic effect of the active ingredient. Solvents may include dimethylformamide, trifluoroacetic acid, dimethyl sulfoxide (DMSO), and dihydro-L-glucanone (CYRENE). TM N-methylpyrrolidone (NMP), halogenated analgesics (selected from halothane, isoflurane, enflurane, desflurane, and sevoflurane), and mixtures and combinations thereof. In one embodiment, the solvent is DMSO.

[0058] In embodiments, the solvent may be applied in an amount sufficient to enhance the therapeutic efficacy of the dicarboxylate of formula I. In some embodiments, the solvent (e.g., DMSO) may be applied in an amount sufficient to reduce the pain response by, for example, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%. In embodiments, the amount of solvent or DMSO applied is sufficient to enhance the efficacy of the dicarboxylate of formula I in lowering the blood glucose level in the subject, thereby treating one or more disorders, diseases, or conditions (e.g., insulin resistance, type 2 diabetes) caused by elevated blood glucose levels. In another embodiment, the amount of solvent or DMSO applied is sufficient to enhance the efficacy of the dicarboxylate of formula I in inhibiting hemolysis caused by the toxin, such that the application can treat disorders, diseases, and conditions associated with PLD toxin.

[0059] In an embodiment, the amount of solvent applied may be sufficient to enhance the therapeutic effect of the dicarboxylic acid ester in providing the above-mentioned effects. The amount of the one or more solvents may be in the range of, for example, about 1% to about 80%, about 5% to about 70%, about 10% to about 50%, about 15% to about 40%, about 20% to about 30%, or any value or range therebetween.

[0060] At least one terpene or related terpene used with the active ingredient dicarboxylic acid ester (preferably diethyl azelate) can be any terpene capable of enhancing the therapeutic effect of the active ingredient. The at least one terpene or related terpene can be a terpene compound having a monocyclic or bicyclic structure, and / or having ten carbon atoms in its chemical formula. The terpenes or terpenoids may be selected from the group consisting of: arnica extract, arnica oil, β-caryophyllene, bisabolol, α-cedrene, carene δ, caryophyllene acetate, caryophyllene, citronellol, cyclomethylene citronellol, decyl acetate, ethyl butyrate, ethyl linalool, cis-3-hexenol, pepper extract from Kalasek, limonene, lime terpenes, D-limonene (also known as limonene), mandarin terpenes, menthol, methyl salicylate, myrcene, nonyl acetate, octyl acetate, orange terpenes, α-pinene and β-pinene, citrus terpenes, α-terpenol, α-terpinene, sesquiterpenes and combinations thereof. In one embodiment, the terpenes are limonene and / or D-limonene, optionally combined with menthol.

[0061] In an embodiment, the amount of the at least one terpene or related terpene applied is sufficient to enhance the efficacy of the dicarboxylic acid ester in providing the aforementioned effects. The amount of the one or more terpenes or terpenoids may range from, for example, from about 0.1% to about 20%, from about 0.25% to about 10%, from about 1% to about 5%, from about 2% to about 4%, or any value or range therebetween. method

[0062] In embodiments, this disclosure provides a method for inducing analgesia to suppress pain in a subject, comprising administering to the subject a pharmaceutical composition comprising an azelaic acid ester, at least one solvent, and at least one terpene. In embodiments, this disclosure provides a method for inducing analgesia to suppress pain in a subject, comprising administering to the subject the aforementioned pharmaceutical composition.

[0063] Local analgesia can be induced by disrupting or increasing the fluidity of lipid rafts. Lipid rafts (also known as lipid microdomains) are discrete lipid domains located in the outer lobules of the plasma membrane. Lipid rafts are rich in cholesterol, sphingomyelin, gangliosides, and sphingolipids, such as monosialotetrahexosylganglioside 1 (GM1). Lipid rafts affect membrane fluidity and membrane protein transport, thereby regulating neurotransmission and receptor transport. [Korade Z, et al., Neuropharmacology (2008) 55(8):1265–73].

[0064] In the implementation scheme, the analgesic effect is induced by the disruption of lipid rafts due to contact between the dicarboxylic acid ester of Formula I and the lipid rafts. When the dicarboxylic acid ester of Formula I is applied topically to a subject (e.g., an open wound, peri-wound area, healthy skin, mucous membrane, or oral mucosal wound), the dicarboxylic acid ester contacts and disrupts the lipid rafts located in the subject's plasma membrane. In the implementation scheme, the lipid rafts contain GM1.

[0065] In one embodiment, the lipid raft contains phospholipase D2 (PLD2). In another embodiment, the dicarboxylic acid ester of Formula I disrupts the lipid raft containing GM1. In yet another embodiment, the disruption of the lipid raft by the dicarboxylic acid ester of Formula I causes PLD2 to leave the lipid raft, thereby activating TREK-1 and subsequently promoting the production of the signaling lipid phosphatidic acid (PA).

[0066] Pain can be subdivided in many ways by type. Based on duration, pain can be classified as chronic or acute. Chronic pain is considered pain that lasts longer than the normal healing time (usually more than 3 months). Chronic pain affects approximately 20% of the world's population and accounts for about 20% of medical visits. [Treede Z, et al., Pain (2015) 156(6): 1003-7]. Based on mechanism, pain can be nociceptive, neuropathic, or nociplastic. Nociceptive pain is caused by mechanical, chemical, or thermal stimuli that damage pain receptors (nociceptors) in tissues. Neuropathic pain is caused by damage to components of the nervous system. Nociplastic pain is caused by alterations in the perception of nociception.

[0067] In this implementation plan, the subject experiences pain. The pain may be acute or chronic. The pain may be nociceptive, neuropathic, or nociplastic.

[0068] In this implementation plan, the pain is acute. Subjects may require treatment for acute pain. Examples of acute pain include, but are not limited to, traumatic pain, surgical (surgical, dental, dermatological, etc.) pain, wound pain, musculoskeletal pain (e.g., back pain, neck pain), toothache, infection (e.g., wound infection), and toxins (insect, animal, bacteria, fungus, etc.).

[0069] In the implementation plan, the pain is chronic. In some implementation plans, the subject may need to be treated for chronic pain. Examples of chronic pain include, but are not limited to, fibromyalgia, arthritis pain, iliotibial band syndrome pain, tennis elbow pain, cancer pain, musculoskeletal pain (e.g., back pain, neck pain), temporomandibular joint disorder, trigeminal neuralgia, chronic headache, pain associated with neurological disorders (MS, diabetic neuropathy, etc.), neuroma, pelvic inflammatory disease, endometriosis, herpes zoster and postherpetic neuralgia, and chronic neuropathy associated with infections (e.g., HIV), chemotherapy-induced neuropathic pain, surgery-induced neuropathic pain, trauma-induced neuropathic pain, vulvar pain, atypical craniofacial pain, sciatica, phantom limb, toothache, and burning mouth syndrome.

[0070] In this implementation, the pain is nociceptive pain. In this implementation, the subject may need to be treated for nociceptive pain. Nociceptive pain is typically acute and occurs in response to a specific condition. Examples of nociceptive pain include, but are not limited to, pain from sprains, burns, bruises, surgery, and fractures. When pain persists for more than six months, certain conditions can lead to chronic nociceptive pain. Examples of chronic nociceptive pain include pain caused by cancer, rheumatoid arthritis, osteoarthritis, and musculoskeletal disorders such as back pain.

[0071] In the implementation scheme, the pain is neuropathic pain. In the implementation scheme, the subject may require treatment for neuropathic pain. Neuropathic pain is caused by damage to components of the nervous system. Certain diseases may be the underlying cause of neuropathic pain. For example, the subject may have metabolic diseases (e.g., diabetic neuropathy), autoimmune diseases (e.g., multiple sclerosis), viral infections (e.g., herpes zoster and its sequelae, postherpetic neuralgia), vascular diseases (e.g., stroke), trauma, and / or cancer. [Campbell JN et al., Neuron (2006) 52(1):77-92; Dworkin RH et al., Arch Neurol (2003) 60; 1524-34]. In the implementation scheme, neuropathic pain is caused by nerve damage due to metabolic diseases, trauma, ischemia or hemorrhage, inflammation, neurotoxicity, neurodegeneration, paraneoplasticity, vitamin deficiency, or cancer. Examples of neuropathic pain include, but are not limited to, postherpetic neuralgia (or post-shingles), reflex sympathetic dystrophy / burning pain (neurotrauma), components of cancer pain, phantom limb pain, entrapment neuropathy (e.g., carpal tunnel syndrome), and peripheral neuropathy (extensive nerve damage).

[0072] In the implementation scheme, the pain is nociplastic pain. In the implementation scheme, the subject may need to be treated for nociplastic pain. Nociplastic pain is pain caused by altered nociceptive sensation, although there is no clear evidence that actual or threatening tissue damage causes activation of peripheral nociceptors, nor is there evidence that disease or lesion of the somatosensory system causes pain. [Chimenti RL et al., PhysTher. (2018) 98(5): 302–314].

[0073] In the implementation plan, pain is selected from acute pain, chronic pain, nociceptive pain, neuropathic pain, nociplastic pain, traumatic pain, chemical pain, burn pain, ischemic pain, insect bite pain, stinging pain, musculoskeletal pain (e.g., back pain, neck pain), rheumatoid arthritis pain, postoperative pain, bone pain (e.g., osteoarthritis), and pain caused by various skin diseases (e.g., acne, psoriasis, hidradenitis suppurativa, eczema, rosacea).

[0074] In the implementation scheme, the reduction in pain (or the reduction in pain response) is due to the regulation of PRR activity. In the implementation scheme, the reduction in pain response (or the decrease in pain response) is due to the regulation of NOD receptor activity. In the implementation scheme, the reduction in pain response (or the decrease in pain response) is due to the regulation of Dectin receptor activity. Synthesis of dicarboxylic acid esters

[0075] Dicarboxylic acid esters of Formula I are commercially available or prepared by various methods known in the art. In embodiments, dicarboxylic acid esters can be prepared by direct ester formation from a desired acid and alcohol. This condensation can be achieved by dehydrating the reaction mixture with a suitable reagent or by heating the mixture of acid and alcohol. In embodiments, dicarboxylic acid esters can be prepared by reacting an alcohol with an activated form of an acid. Activated forms of acids include acyl halides, acid anhydrides including homoanhydrides and heteroanhydrides, reactants of the parent acid's anhydride with a desired alcohol, esters of both the acid and alcohol, and acid anhydrides, which are formed by reacting the desired acid or alcohol with p-toluenesulfonyl chloride to produce toluenesulfonyl anhydride or ester, which is subsequently reacted with an alcohol or acid, respectively, to produce the desired final ester. Similarly, simple organic acid anhydrides (e.g., acetic anhydride) can be used instead of p-toluenesulfonyl chloride. Alternatively, the starting ester of the acid can be converted to an ester by dissolving the ester in the desired alcohol in the presence of a suitable acidic or basic catalyst, wherein the alcohol becomes the reactant; this method is also known in the art as transesterification.

[0076] For example, a dimethyl ester of an acid can be started, and a diethyl ester of that acid can be readily formed by dissolving the ester in ethanol in the presence of an acid or base. Alternatively, if a mixed ester of the acid is desired, a solution of two or more desired alcohols of appropriate composition can be used in any of the methods described herein.

[0077] The desired ester can be formed by using halogenation intermediates or components. For example, thionyl chloride will chlorinate an acid and an alcohol to produce an acyl chloride and an alkyl chloride. These acyl chlorides and alkyl chlorides can then be further reacted with the desired alcohol or acid, respectively, to produce the desired ester product. Other common halogenating agents include, for example, oxalyl chloride, as well as chlorides and bromides of phosphorus, such as phosphorus pentachloride or phosphorus trichloride, and phosphorus pentabromide or phosphorus tribromide or phosphorus oxychloride.

[0078] The common practice is to form esters by the action of a strong base on a mixture of acid and alcohol. Examples of strong bases include lithium aluminum hydride and other metal hydrides, alkali metal alkoxides (such as sodium ethoxide), and diisobutylaluminum hydride. Application method

[0079] The pharmaceutical compositions disclosed herein can be administered to a subject in a variety of ways. For example, the pharmaceutical compositions can be administered topically, transdermally, intravenously, subcutaneously, intramuscularly, or orally. The pharmaceutical compositions can be applied topically to the target area or the area requiring treatment and / or around it.

[0080] Treatment regimens can vary based on a variety of factors that are typically considered by those skilled in the art. These factors include the route of administration, the nature of the formulation, the nature of the patient's disease, the subject's body type, weight, surface area, age, sex, other medications administered to the patient, and the judgment of the attending physician. The pharmaceutical composition may be administered concurrently with or as a supplement to other treatments for inflammatory conditions or pain relief.

[0081] The pharmaceutical composition may be administered in combination with one or more other therapeutic agents to treat pain and / or inflammation. These other therapeutic agents may be administered via the same or different routes of administration. These other therapeutic agents may include analgesics, anti-inflammatory agents, anesthetics, antibiotics, and antifungals.

[0082] In one embodiment, the pharmaceutical composition is applied topically to the subject's skin, for example, at or near the area requiring treatment. In another embodiment, the pharmaceutical composition is administered to the subject by rubbing it onto the skin, allowing the composition (or at least the dicarboxylic acid ester) to be absorbed through the skin.

[0083] In the implementation scheme, for prophylactic treatment, the pharmaceutical composition may be administered to the subject within any range of time intervals from which it can be derived: 96 hours to immediately prior to surgery, 72 hours to immediately prior to surgery, 48 hours to immediately prior to surgery, 24 hours to immediately prior to surgery, 12 hours to immediately prior to surgery, 8 hours to immediately prior to surgery, 4 hours to immediately prior to surgery, 2 hours to immediately prior to surgery, 1 hour to immediately prior to surgery, or 0.5 hours to immediately prior to surgery or immediately prior to surgery. The pharmaceutical composition may be applied topically to a healthy skin area at any time, for example, in one implementation scheme, before hiking.

[0084] In the implementation plan, for the purpose of preventive treatment, the pharmaceutical composition is applied locally or topically to the subject prior to surgery such as venipuncture, injection, incision, hair removal, tattoo application and removal.

[0085] In the implementation plan, the pharmaceutical composition is applied locally or topically to the subject during or after procedures such as venipuncture, injection, incision, hair removal, tattoo application and removal.

[0086] In the implementation plan, the pharmaceutical composition is administered to the subject once or multiple times. For example, the pharmaceutical composition may be administered at predetermined intervals. In the implementation plan, for example, the pharmaceutical composition may be administered once daily, twice daily, three times daily, four times daily, or more than four times daily, or every other day, every three days, every four days, etc.

[0087] In the implementation plan, the pharmaceutical composition is administered to the subject at a therapeutically effective dose. The therapeutically effective dose, when administered to the subject, will depend on the specific condition being treated and the desired outcome.

[0088] The pharmaceutical composition may be administered to a subject in need approximately every 1 to 24 hours, approximately every 1 to 12 hours, approximately every 2 to 8 hours, approximately every 2 to 6 hours, approximately every 4 hours to 6 hours, approximately every 4 to 8 hours, approximately every 12 hours, approximately every 24 hours, approximately every 48 hours, or more frequently. In embodiments, the pharmaceutical composition may be administered once, twice, three times, four times, five times, six times, seven times, eight times, or more frequently, or any combination thereof. In embodiments, the pharmaceutical composition may also be administered daily, every other day, every two days, every three days, every four days, or less frequently, or any combination thereof. In embodiments, the pharmaceutical composition may be administered to a subject in need for a duration of 1 to 30 days, 1 to 25 days, 1 to 20 days, 1 to 15 days, or 1 to 10 days. formula

[0089] In embodiments, the pharmaceutical compositions disclosed herein can be formulated for delivery via any route of administration known in the art, including but not limited to topical, transdermal, intravenous, subcutaneous, intramuscular, or oral administration.

[0090] Appropriate formulations depend on the chosen route of administration. Any well-known techniques, carriers, and excipients as understood in the art may be used, such as those disclosed in Remington's Pharmaceutical Sciences, 18th ed. (Mack Publishing Company: Easton, PA., 1990), which is incorporated herein by reference. The pharmaceutical compositions disclosed herein may be manufactured in any manner known in the art, such as by conventional methods of mixing, dissolving, granulating, grinding, emulsifying, encapsulating, embedding, or compressing.

[0091] Pharmaceutical compositions include those suitable for topical, transdermal, intravenous, subcutaneous, intramuscular, or oral administration, although the most appropriate route may depend, for example, on the recipient's condition and illness.

[0092] In embodiments, the pharmaceutical compositions are suitable for topical and transdermal application. Topical and transdermal application of the azelaic esters disclosed herein can be in the form of processed gels, creams, lotions, solutions, ointments, suspensions, or emulsions. These pharmaceutical compositions may also contain one or more suitable excipients disclosed herein.

[0093] Parenteral administration (e.g., intravenous, subcutaneous, or intramuscular) of the dicarboxylic acid esters of Formula I can be in the form of a solution, suspension, or emulsion. In one embodiment, these formulations are prepared in an aqueous saline solution. These pharmaceutical compositions may also contain one or more suitable excipients disclosed herein.

[0094] In the embodiments, the pharmaceutical composition is suitable for oral administration. The pharmaceutical composition can be conveniently present in unit dose form and can be prepared by any method known in the pharmaceutical field. Typically, these methods involve mixing a dicarboxylic acid ester of Formula I and optionally any co-administered active ingredient with a carrier constituting one or more excipients. Generally, the pharmaceutical composition is prepared by uniformly and tightly mixing the active ingredient with a liquid carrier or a finely chopped solid carrier (or both), and then shaping the product into the desired composition as needed. Pharmaceutical compositions suitable for oral administration and any optional second active ingredient can be presented as: discrete units, such as capsules, pouches, or tablets, each unit containing a predetermined amount of active ingredient; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; or oil-in-water or water-in-oil liquid emulsions. The active ingredient can also be present as pills, saccharides, or pastes. These pharmaceutical compositions may also contain one or more suitable excipients disclosed herein. excipient

[0095] In embodiments, the pharmaceutical compositions of this disclosure may further comprise one or more excipients. Excipients may include carriers, such as water-insoluble polysaccharides or oligosaccharides. Examples of carriers include, but are not limited to, cellulose acetate, cellulose acetate butyrate, cellulose acetate propionate, cellulose acetate phthalate, chitosan, β-cyclodextrin, ethyl cellulose, hydroxypropyl methyl phthalate (HPMCP), microcrystalline cellulose, starch, and any combination thereof.

[0096] Excipients may include thickeners, such as water-soluble polysaccharides. Examples of thickeners include, but are not limited to, hydroxypropyl methylcellulose (HPMC), gum arabic, alginate, colloidal silica, calcium carboxymethyl cellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose (hydroxypropyl methylcellulose), methylcellulose, sucrose, sodium alginate, sodium carboxymethyl cellulose, and any combination thereof.

[0097] In embodiments, the pharmaceutical compositions of this disclosure may further comprise one or more pharmaceutical excipients, such as ascorbic acid, EDTA dihydrate, glycerin, citrate monohydrate, sodium citrate dihydrate, sodium benzoate, sodium propionate, 70% sorbitol solution, sucralose, FD&C Yellow #6, artificial flavorings (e.g., artificial mint flavoring, artificial fruit flavoring), purified water, or any combination thereof.

[0098] In embodiments, the pharmaceutical compositions of this disclosure may comprise preservatives. Suitable preservatives include, but are not limited to, mercury-containing substances, such as phenylmercuric salts (e.g., phenylmercuric acetate, borate, and nitrate) and thimerosal; stable chlorine dioxide; quaternary ammonium compounds, such as benzalkonium chloride, hexadecyltrimethylammonium bromide, and hexadecylpyridine chloride; imidazolidinyl urea; parabens, such as methylparaben, ethylparaben, propylparaben, and butylparaben and their salts; phenoxyethanol; chlorophenoxyethanol; phenoxypropanol; chlorotert-butanol; chlorocresol; phenethyl alcohol; disodium ethylenediaminetetraacetate; benzoic acid, benzyl alcohol, and sorbic acid and their salts.

[0099] In embodiments, the pharmaceutical compositions of this disclosure may comprise one or more acceptable pH adjusters and / or buffers, including acids such as acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and tris(hydroxymethyl)aminomethane; and buffers such as citrate / glucose, sodium bicarbonate, and ammonium chloride. The amounts of such acids, bases, and buffers are sufficient to maintain the pH of the composition within a pharmaceutically acceptable range. In embodiments, the pH of the pharmaceutical compositions of this disclosure may be from pH 4 to pH 7.5.

[0100] In embodiments, the pharmaceutical compositions of this disclosure may comprise sugar alcohols. Examples of sugar alcohols include, but are not limited to, mannitol, glycerol, galactitol, fucitol, inositol, vortexitol, maltotriol, maltotetratitol, polyethylene glycol, erythritol, threitol, ribitol, arabinitol, xylitol, allitol, galactitol, glucositol, sorbitol, arabinitol, maltitol, lactitol, isomaltitol, and any combination thereof.

[0101] In embodiments, the pharmaceutical compositions of this disclosure may include additives. Examples of additives include, but are not limited to, diluents, binders, surfactants, lubricants, flow aids, coating materials, plasticizers, colorants, flavoring agents, or pharmaceutically inert materials. Examples of diluents include, for example, cellulose; cellulose derivatives such as microcrystalline cellulose; starch; starch derivatives such as corn starch, cyclodextrin, etc.; sugars; sugar alcohols such as lactose, D-mannitol, etc.; inorganic diluents such as dried aluminum hydroxide gel, precipitated calcium carbonate, magnesium aluminum metasilicate, calcium hydrogen phosphate, etc. Examples of binders include, for example, hydroxypropyl cellulose, methylcellulose, hydroxypropyl methylcellulose (hydroxypropyl methylcellulose), povidone, dextrin, pullulan, hydroxypropyl starch, polyvinyl alcohol, alginate, agar, gelatin, tragacanth gum, polyethylene glycol, etc. Examples of surfactants include, for example, sucrose esters of fatty acids, polyoxyethylene stearates, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene glycol, sorbitol sesquioleate, sorbitol trioleate, sorbitol monostearate, sorbitol monopalmitate, sorbitol monolaurate, polysorbate, glyceryl monostearate, sodium lauryl sulfate, polylauryl alcohol, and quaternary ammonium salts (e.g., benzyl dimethyltetradecyl ammonium chloride hydrate, benzyl chloride, benzyl hexadecyl dimethyl ammonium chloride hydrate, benzyl dimethyl stearyl ammonium chloride hydrate, benzyl dodecyl dimethyl ammonium chloride dihydrate, benzyl dodecyl dimethyl ammonium bromide). Examples of lubricants include, for example, stearic acid, calcium stearate, magnesium stearate, and talc. Examples of flow aids include, for example, dried aluminum hydroxide gel and magnesium silicate. Examples of coating materials include, for example, hydroxypropyl methylcellulose 2910, aminoalkyl methacrylate copolymer E, polyvinyl acetal diethylaminoacetate, polyethylene glycol 6000, and titanium dioxide. Examples of plasticizers include, for example, triethyl citrate, triacetin, and polyethylene glycol 6000. Reagent test kit

[0102] In the embodiments, this disclosure also provides a kit comprising the pharmaceutical composition of this disclosure (especially for topical application), a wiping agent for cleaning the site, a swab or brush for applying the applied material, and an adhesive dressing for covering the site; optionally, the kit may include a disposal bag or container; optionally, the kit may include an absorbent element; optionally, the kit may include a pair of gloves (e.g., sterile nitrile or latex gloves). Example Materials and Methods

[0103] Chemicals: Azelaic acid diester was synthesized from azelaic acid and ethanol via acid-catalyzed esterification, followed by fractionation to a purity of 99% and determination by gas chromatography-mass spectrometry (GC-MS), as described in Izbicka E, Streeter Rand Louden C: Adaptive membrane fluidity modulation: A feedback regulated homeostatic system and target for pharmacological intervention In Vivo 35 (3073-3095, 2021). Unless otherwise stated, all chemicals are from Sigma-Sigma-Aldrich (St. Louis, MO) and Thermo Fisher Scientific (Waltham, MA). Terpenes are from Vigon International (East Stroudsburg, PA).

[0104] The tactile sensory assessment kit (North Coast Medical Inc, Morgan Hill, CA) contains 20 monofilaments that apply target forces between 0.25 and 512 millinewtons (mN), corresponding to 0.008 and 300 g, respectively. The applied forces are numerically coded in milligrams (mg) as follows: 1.65, 2.36, 2.44, 2.83 (normal sensation), 3.22, 3.61, 3.84 (weakened light touch), 4.08, 4.17, 4.31 (weakened protective sensation), and 4.56, 4.74, 4.93, 5.07, 5.18, 5.46, 5.88, 6.10, 6.45, 6.65 (loss of protective sensation). Notably, the logarithmic transformation makes the variance (dispersion) more uniform across the different treatment groups. See Mills C, et al., "Estimating efficacy and drug ed50's using von frey thresholds: Impact of weber's law and logtransformation," J Pain 13(6):519-523, 2012.

[0105] The CMS (Care-Sensitive Skin Test) uses a monofilament to test sensory levels and obtain data on sensory attenuation or recovery, following modified protocols from the following literature: Rolke R et al., “Quantitative sensory testing: A comprehensive protocol for clinical trials,” Eur J Pain 10(1):77-88, 2006, and Kostek M, et al., “A protocol of manual tests to measure sensation and pain in humans,” J Vis Exp 118), 2016. The tests were performed at approximately the same time in the morning. Subjects lay face down on a flat surface, and circular test areas with a radius of 10 cm were marked on the back of the left and right popliteal fossa areas behind the knees. To establish a baseline of the minimum force required to generate tactile sensation, these areas were randomly touched with a monofilament, and a series of ten measurements (five on each side) were performed, each measurement using a monofilament with increasing force. Each touch was recorded, and if the subject felt tactile sensation, they reported it. A positive score for a monofilament was defined as ≥6 reported tactile events.

[0106] Test sample solutions were prepared in isopropanol. To test DEA and DMSO at their highest equimolar concentrations of approximately 3 M each, pure stock solutions without diluent were prepared and adjusted according to the densities of the two compounds to correspond to 78% (v / v) DEA and 22% (v / v) DMSO. These compounds were evaluated using single reagents and equimolar stock solutions at different proportions (DEA concentrations of 78%, 58%, 39%, and 19% (v / v), and DMSO concentrations of 22%, 16%, 11%, and 5% (v / v), rounded to the nearest integer). After adding other components, the highest concentrations of DEA and DMSO were 77% and 21%, respectively.

[0107] After baseline assessment, 0.5 ml of test sample solution was applied to each test area and gently rubbed for 15 seconds (Day 1, time = 0). Mechanosensitivity testing was then performed at the earliest 5 minutes (0.1 hours), followed by mechanosensitivity testing at 1, 4, 8, 24, 48, 72, and 96 hours. The test was completed when tactile sensation returned to baseline, and this period was further referred to as the time to baseline (TBL). Unless otherwise specified, all tests were performed in 10 replicates.

[0108] The area under the curve (AUC) for each test cycle from t=0 to recovery to baseline sensitivity was determined using SigmaPlot version 14.5 (Inpixion, Palo Alto, CA). Statistical analysis was performed using the Student's t-test; a p-value < 0.05 was considered significant, and a p-value < 0.005 was considered highly significant.

[0109] Hemolysis tests were performed in peripheral human blood using a bee venom preparation containing the active hemolytic component PLA2, repeated three times, as described in Streeter RT and Izbicka E, “Diethyl azelate for the treatment of brownrecluse spider bite, a neglected orphan indication,” In Vivo 36(1):86-93, 2022. In short, the bee venom stock solution was diluted in phosphate-buffered saline (PBS) containing 0.5% Tween (diluent) to achieve a level of hemolysis comparable to that of Triton-X used as a positive control. All test samples were also prepared in the same diluent. The reagents were pre-incubated at room temperature for 5 minutes, followed by a 30-minute reaction initiated by the addition of a red blood cell suspension.

[0110] To assess the effects of DEA alone and a mixture of DEA, DMSO, and limonene (77%, 21%, and 2%, respectively) on blood glucose, 12.5 μL (approximately 12.5 mg) of the solution was administered orally. Blood glucose levels were quantified every 15 minutes before administration and for 2 hours thereafter using a OneTouch Ultra2 blood glucose meter (LifeScan, Malvern, PA) and UniStrip1 test strips (UniStrip Technologies, Charlotte, NC) according to the manufacturer's instructions (8 measurements were repeated per treatment). Example 1 - Evaluation of DEA and DMSO in Skin Mechanosensitivity Test

[0111] This experiment demonstrates that the combined use of DEA and DMSO exhibits an unexpected synergistic effect and nonlinear concentration dependence in the skin mechanosensitivity test (CMS).

[0112] The CMS test (n=20) determined that the baseline sensitivity without any treatment was 2.36 monofilaments and 0.02 g bending strength. DEA and DMSO were tested, either as single reagents or as a mixture of DEA and DMSO. The results summarized in Table 1 (where a relative molar ratio of 1 corresponds to 78% DEA and 22% DMSO) show that an increase in the molar ratio was not associated with an increase in the time to baseline (TBL). This unusual relationship can be understood by comparing the data for DEA and DMSO, especially at molar ratios above 0.75. Table 1 below shows the relationship between the relative molar concentrations (Rm) of DEA and DMSO and the time to baseline in the skin mechanosensitivity test. Table 1

[0113] Table 2 provides more extensive data in the DEA and DMSO concentration matrix. For mixtures of both compounds, the endpoint is expressed as time to baseline (TBL), with a correction for the contribution of DMSO to the mixture after subtracting the effect of DEA alone from the TBL of the mixture (TBL-C). TBL values ​​are uniformly repeatable, therefore no standard deviation is shown. AUC values ​​and standard deviations are shown. The most significant effect was observed at a TBL of 72 hours (TBL-C of 68), at which the percentages of DEA and DMSO were 78 / 22 and 58 / 22, respectively. The latter test used fibers of 4.74, 4.03, and 5.07, which applied flexural forces of 6–10 g. The results indicated a significant loss of protective sensation compared to the untreated control group. Table 2 below shows the relationship between DEA and DMSO concentrations, AUC, and time to baseline in the skin mechanosensitivity test. Table 2

[0114] Figure 1 The unexpected properties of the combination of DMSO and DEA are highlighted. DMSO concentrations below 10% are ineffective, but the analgesic effect is dramatically enhanced to a maximum at 22% DMSO and 78% DEA, subsequently decreasing sharply at lower DEA concentrations. These data suggest that the enhanced analgesic effect of DEA is not solely attributable to the increased tissue penetration of DMSO. Example 2 - Screening of local analgesics, turpentine oil and terpenes

[0115] These experiments demonstrate that certain combinations of turpentine, limonene, pinene, and menthol with DEA ​​and DMSO exhibit unexpectedly superior activity.

[0116] The initial CMS test was performed using selected local analgesics. The time to baseline (TBL) values ​​for 4% lidocaine and diclofenac (1.16% diclofenac) were 4 hours and 8 hours, respectively. Turpentine oil (also known as terpenes) was also tested in the CMS screening because it is a component of some analgesics. Compared to the carrier control, mixtures of 1% and 2% turpentine oil with 21% DEA and 77% DMSO produced TBL values ​​ranging from 24 to 48 hours.

[0117] Turpentine is a highly variable mixture. Products manufactured in the United States typically contain α-pinene (75% to 85%), varying amounts of α-pinene (up to 3%), camphene (4% to 15%), limonene (5% to 15%), and low amounts of 3-carene and terpinene.

[0118] To assess the relative contribution of turpentine components in CMS testing, screening was performed using a mixture of 19% DEA and 5% DMSO, along with the following turpentine components and associated terpenes. All compounds were tested individually at 2% v / v: pentanol, arnica extract, arnica oil, β-caryophyllene, bisabolol, α-cedrene, carene δ, caryophyllene acetate, caryophyllene, citronellol, cyclomethylene citronellol, decyl acetate, ethyl butyrate, ethyl linalool, cis-3-hexenol, pepper extract from Karsek, limonene, sour orange terpenes, D-limonene (also known as limonene), citrus terpenes, menthol, methyl salicylate, myrcene, nonyl acetate, octyl acetate, orange terpenes, α-pinene and β-pinene, citrus terpenes, α-terpenol, α-terpinene and valencene. In addition, cannabinoid oil (30%) and synthetic capsaicin nonanoic acid vanillamide (PAVA) (0.25% to 4%) were also detected. In all cases, the time-to-batch (TBL) of the detected compounds was at least 1 hour, but except for menthol, α-pinene, and D-limonene, there were no statistically significant differences between the treatment and control groups. These compounds will be further evaluated in Example 3 below. Example 3 - Evaluation of multi-component mixtures in CMS determination

[0119] This example illustrates that certain combinations of DEA, DMSO, menthol, α-pinene, and limonene unexpectedly enhance TBL. Furthermore, in diesters of azelaic, octanoic, and sebacic acids, only DEA ± limonene significantly inhibited CMS, regardless of the presence of limonene.

[0120] Table 3 summarizes the effects of different concentrations of DEA and DMSO combined with limonene. The last column shows the TBL values ​​(TBL-C) after correction for the contributions of DEA and DMSO (without limonene). Bold text in TBL-C indicates values ​​higher than the contributions of DEA and DMSO. Table 3 below shows the relationship between DEA, DMSO, and limonene concentrations, AUC, and time to baseline in the skin mechanosensitivity test. Table 3

[0121] As shown in Table 3, the TBL-C was longest at 20 hours for equimolar DEA and DMSO (19% and 5%, respectively) when 2% limonene was used. The TBL-C values ​​for equimolar DEA and DMSO (39% and 11%, respectively) decreased to 16 hours. However, 2% limonene did not further enhance the combined activity observed at their respective highest concentrations (77% and 21%, respectively) for equimolar DEA and DMSO.

[0122] It is noteworthy that 2% limonene has a synergistic effect with lower percentages of DEA and DMSO. As shown in Tables 2 and 3, the TBL-C of the 39% DEA and 11% DMSO mixture is 7 hours without limonene, but 16 hours with 1% or 2% limonene.

[0123] The dose-response of limonene was evaluated using 20% ​​DEA and 7% DMSO. Figure 2 A and Figure 2 B). After adjusting for contributions from DEA and DMSO, the highest TBL values ​​for 4% and 2% limonene were 16 hours. 10% limonene had no effect, while 0.25% limonene reduced the TBL to below the levels seen with DEA ​​plus DMSO. Limonene also exhibited a bell-shaped dose-response curve.

[0124] Given that 2% limonene significantly enhances TBL in a DEA / DMSO mixture, other medium-chain fatty acid diesters at a concentration of 39% were also tested in 11% DMSO ± 2% limonene. Figure 3 A and Figure 3 As shown in B, DEA was superior to other diesters, and limonene significantly enhanced the effect of DEA. Similarly, limonene exhibited peak analgesic activity between 1% and 2%, but the contribution of limonene decreased at higher concentrations and did not provide additional benefit above 4%. Figure 2 A and Figure 2 B).

[0125] In a head-to-head comparison of pinene isomers (Table 4), 2% α-pinene was significantly superior to β-pinene in DEA / DMSO mixtures of 39% / 11% and 19% / 5%. At the same molar concentration in DEA / DMSO, the activity of 2% α-pinene was comparable to that of 2% limonene (see Table 2). However, the analgesic effect of mixtures of 2% limonene and 2% α-pinene in DEA / DMSO was not significant. Table 4 below shows the relationships between the concentrations, AUC, and time to baseline of DEA, DMSO, limonene, and pinene in CMS assays. Table 4

[0126] As shown in Table 5, menthol (0.1% to 4%) was ineffective when mixed with 39% DEA. Adding 0.1% menthol to DEA / DMSO mixtures in 39 / 11 and 19 / 5 percentage ratios resulted in a surprising enhancement of analgesic effects. 0.1% menthol significantly enhanced the analgesic effects of DEA / DMSO / limonene at 39 / 11 / 2 and 19 / 5 / 2 ratios. The observed maximum TBL for DEA / DMSO / limonene / menthol (39 / 11 / 2 / 0.1%) was 48 hours, significantly higher than that of the DEA / DMSO / limonene mixture without menthol (TBL-C was 16 hours; see Table 3). Table 5 below shows the effect of menthol on the duration of inhibition of sensitivity in a mixture of DEA, DMSO, and limonene in a skin mechanosensitivity test. Table 5

[0127] Figure 4 The effects of 39% DEA, 11% DMSO, 2% limonene, and 0.1% menthol as single agents and in combination on the duration of sensitivity inhibition in CMS assays were summarized. Both the four-component and three-component mixtures showed superior activity compared to any single agent or two-component mixture. Example 4 - Comparison of the effects of oral DEA and a mixture of oral DEA, DMSO and limonene on blood glucose levels

[0128] This experiment compared the effects of oral DEA and a mixture of oral DEA, DMSO, and limonene on blood glucose levels.

[0129] Figure 5 The figures show blood glucose levels following a single oral dose of 12.5 μL DEA (equivalent to 0.17 mg / kg DEA) (top dashed line) and blood glucose levels following a single oral dose of a mixture of 12.5 μL 77% DEA, 21% DMSO, and 2% limonene (equivalent to 0.13 mg / kg DEA) (bottom solid line). The mean difference in mean blood glucose levels between the two formulations at 2 hours was 39%, which was highly significant (p = 0.001).

[0130] Previous studies have shown that oral DEA at a dose of 1 mg / kg provides optimal glycemic inhibition. Current results indicate that a mixture of 77% DEA, 21% DMSO, and 2% limonene is approximately 5.8 times more potent than DEA alone. Therefore, embodiments include a composition containing approximately 70% to approximately 80% DEA, approximately 19% to approximately 22% DMSO, and approximately 1% to approximately 3% limonene, or 76%–78% DEA, 20.5% to 21.5% DMSO, and 0.5% to 1.5% limonene. Example 5 - Composition Evaluation of PLA2-Induced Hemolysis Mixture

[0131] This example demonstrates that, compared to a single reagent, the mixture of DEA, DMSO, and limonene exhibits an unusual bell-shaped dose and concentration effect on the degree of PLA2-induced hemolysis in bee venom.

[0132] Figure 6 In the figures, columns 1-5 show the bell-shaped hemolytic inhibitory activity of the mixture of DEA, DMSO, and limonene. Notably, the concentration at which the maximum inhibitory effect occurred was 50 times lower than the effective concentration in CMS. At the concentration corresponding to the maximum inhibitory effect, individual components were ineffective in this assay (columns 6-9). Furthermore, a comparison of the three diethyl esters (C-8 octanoate, C-9 azelaate, and C-10 sebacic acid ester; all at 5%) showed that diethyl sebacic acid significantly inhibited hemolysis by 60%.

[0133] Although these embodiments have been described with reference to particularly preferred features, examples and preferred embodiments, those skilled in the art will understand that various modifications can be made to these preferred embodiments without departing from the spirit and scope of the invention.

Claims

1. A pharmaceutical composition comprising: Dicarboxylic acid esters of Formula I: R2OOC-(CH2) n -COOR1 Where n is between 4 and 10, and R1 and R2 are each independently C1 to C6 saturated straight-chain or branched alkyl groups; At least one solvent; and At least one terpene.

2. The pharmaceutical composition according to claim 1, wherein the dicarboxylic acid ester of formula I is azelaic acid ester, wherein the azelaic acid ester is selected from diethyl azelaate, diisopropyl azelaate, and mixtures thereof.

3. The pharmaceutical composition according to claim 2, wherein the azelaic acid ester is diethyl azelaic acid ester.

4. The pharmaceutical composition according to claim 1, wherein the content of the dicarboxylic acid ester of formula I ranges from about 30% to about 90% by weight of the composition.

5. The pharmaceutical composition according to claim 1, wherein the at least one solvent is selected from the group consisting of: dimethylformamide, dimethyl sulfoxide (DMSO), dihydroglucosamine (CYRENE). TM ), N-methylpyrrolidone (NMP), fluorocarbons, isofluranes, enflurane, desflurane, sevoflurane, and mixtures and combinations thereof.

6. The pharmaceutical composition according to claim 5, wherein the solvent is DMSO.

7. The pharmaceutical composition according to claim 1, wherein the content of the at least one solvent ranges from about 10% to about 50% by weight of the composition.

8. The pharmaceutical composition according to claim 1, wherein the at least one terpene is a terpene or terpene compound selected from the group consisting of: arnica extract, arnica oil, β-caryophyllene, bisabolol, α-cedrene, carene δ, caryophyllene acetate, caryophyllene, citronellol, cyclomethylene citronellol, decyl acetate, ethyl butyrate, ethyl linalool, cis-3-hexenol, pepper extract from Kalasek, limonene, sour orange terpene, D-limonene (also known as limonene), citrus terpene, menthol, methyl salicylate, myrcene, nonyl acetate, octyl acetate, orange terpene, α-pinene and β-pinene, citrus terpene, α-terpenol, α-terpinene, sesquiterpenes and mixtures thereof.

9. The pharmaceutical composition according to claim 8, wherein the terpene is D-limonene.

10. The pharmaceutical composition according to claim 1, wherein the content of the at least one terpene ranges from about 1% to about 5% by weight of the composition.

11. A pharmaceutical composition comprising: Diethyl azelaate comprising approximately 70% to approximately 80%; Approximately 19% to approximately 22% of DMSO; and Limonene, approximately 1% to approximately 3%.

12. A method for inducing analgesic effects and inhibiting pain in a subject, comprising administering to the subject a therapeutically effective amount of a composition, the composition comprising: Dicarboxylic acid esters of Formula I: R2OOC-(CH2) n -COOR1 Where n is between 4 and 10, and each R1 and R2 is independently a C1 to C6 saturated straight-chain or branched alkyl group; At least one solvent; and At least one terpene.

13. The method according to claim 12, wherein the dicarboxylic acid ester of formula I is diethyl azelaate.

14. The method according to claim 12, wherein the content of the dicarboxylic acid ester of formula I ranges from about 30% to about 90% by weight of the composition.

15. The method according to claim 12, wherein the at least one solvent is DMSO.

16. The method of claim 12, wherein the content of the at least one solvent ranges from about 10% to about 50% of the weight of the composition.

17. The method according to claim 12, wherein the at least one terpene is limonene.

18. The method of claim 12, wherein the content of the at least one terpene ranges from about 1% to about 5% by weight of the composition.

19. A method for inducing analgesic effects and inhibiting pain in a subject, comprising administering to the subject a therapeutically effective amount of a composition, the composition comprising: Diethyl azelaate comprising approximately 70% to approximately 80%; Approximately 19% to approximately 22% of DMSO; and Limonene, approximately 1% to approximately 3%.

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