Triphenylphosphonium-bonded salicylamine derivative

Abstract

Novel salicylamine derivatives can directly target mitochondria, increasing efficacy and reducing required dosages in the treatment of conditions caused by inflammation or oxidative stress.

Description

Technical Field

[0001] Cross - References to Related Applications This application claims the benefit of U.S. Provisional Application No. 62 / 796,999, filed on Jan. 25, 2019, the entire disclosure of which is hereby expressly incorporated by reference herein.

[0002] Technical Field The present disclosure generally relates to salicylamine derivatives that can target mitochondria.

Background Art

[0003] Oxidative stress plays an important role as an etiology of many neurodegenerative and cardiovascular diseases. Inflammatory processes are associated with many of the same diseases. It is well known that leukocytes are mobilized to the site of inflammation. Although it is well established that the activation of these inflammatory cells up - regulates the expression of COX - 2 and promotes prostaglandin formation, this is also accompanied by the production of reactive oxygen species (ROS) by these cells via, for example, NADPH oxidase. Thus, oxidative stress is recognized as an important component of inflammation, but conversely, oxidative stress is also causally related to the development of inflammation. Therefore, controlling cellular reactive oxygen species is an important element in the treatment of conditions resulting from inflammation and oxidative stress.

Summary of the Invention

[0004] Some aspects of the present disclosure include classes of lipophilic analogs of pyridoxamine (PM), such as salicylamine (SA, also known as 2 - hydroxybenzylamine or 2 - HOBA), for example, or derivatives of salicylamine.

[0005] In some aspects of the present disclosure, SA and / or derivatives of SA are targeted to mitochondria.

[0006] Some aspects include that SA derivatives conjugated to TPP salts are disclosed herein. To further investigate the potential provided by directly targeting mitochondria, different TPP conjugates of SA are synthesized as further disclosed below.

[0007] Some aspects include a therapeutic dose of the SA-TPP conjugate in the range of at least about 5 μg to about 1000 mg per 24 hours. In some aspects of the present disclosure, the dose of the SA-TPP conjugate administered to a subject within a given 24-hour or equivalent time frame is in the following ranges: at least about 5 μg to about 10 μg, at least about 10 μg to about 20 μg, at least about 20 μg to about 40 μg, at least about 40 μg to about 60 μg, at least about 60 μg to about 80 μg, at least about 80 μg to about 100 μg, at least about 100 μg to about 120 μg, at least about 120 μg to about 140 μg, at least about 140 μg to about 160 μg, at least about 160 μg to about 180 μg, at least about 180 μg to about 200 μg. In some aspects, the dose for a given 24-hour period is from about 200 μg to about 300 μg, or at least about 300 μg to about 400 μg, or at least about 400 μg to about 500 μg, or at least about 500 μg to about 600 μg, or at least about 600 μg to about 700 μg, or at least about 700 μg to about 800 μg, or at least about 800 μg to about 900 μg, or at least about 600 μg to about 700 μg, or at least about 800 μg to about 900 μg, or at least about 900 μg to about 1000 μg. In other aspects, the dose is 1 mg or more per 24 hours, and in some aspects, the dose per 24 hours is at least about 1 mg to about 10 mg, or about 10 mg to about 25 mg, or about 25 mg to about 50 mg, or about 50 mg to about 75 mg, or about 75 mg to about 100 mg.

[0008] Some embodiments include a therapeutic dose of the SA-TPP conjugate in the range of 200 μg to 100 mg every 24 hours. In some aspects of the present disclosure, the dose of the SA-TPP conjugate is 5.0 mg every 24 hours. In some aspects, the dose is 10.0 mg every 24 hours. In other aspects, the dose is 20.0 mg every 24 hours. In still other aspects, the dose is 33.0 mg every 24 hours. In other aspects, the dose is 55.0 mg every 24 hours. In still other aspects, the dose is 82.5 mg every 24 hours.

[0009] The first embodiment of the present disclosure includes a compound of formula I:

[0010]

Chemical formula

[0011] In formula I, R 1 is hydrogen which may have a counterion, R 2 is selected from hydrogen or an alkyl group optionally substituted with R 4 , and any ring position (e.g., ortho, meta or para to the functional group) may be optionally substituted with R 3 is either hydrogen, a hydroxy group, an acyl group, or an alkoxy group, and may be substituted with R 4 . R 4 is a cation which may have a counterion, and n is 1 or 2. The functional group of R 3 may or may not have an intervening alkyl group chain. Any substitution pattern of formula 1 is as follows. R 1 is hydrogen which may have a counterion. R 2 is selected from hydrogen or an alkyl group optionally substituted with R 4 . R 3is a C1-C6 alkyl group optionally substituted with hydrogen, halogen, hydroxy group, acyl group, alkoxy group, alkyl group, alkenyl group, hydroxy group, acyl group or alkoxy group, hydrogen, hydroxy group, acyl group, alkoxy group, hydrogen, hydroxy group or a C1-C6 alkyl group optionally substituted with a C1-C6 alkyl group, a carbonyl group optionally substituted with O, N or S. R 4 is a cation which may have a counter ion, and n is 1 or 2.

[0012] The second embodiment includes a compound of formula I wherein R 3 is hydrogen and R 2 is an alkyl group optionally substituted with R 4

[0013] The third embodiment includes a compound of any one of the first and second embodiments wherein R 4 is a triphenylphosphonium cation.

[0014] The fourth embodiment includes a compound of any one of the first to third embodiments wherein the counter ion is bromide.

[0015] The fifth embodiment includes a compound of any one of the first to fourth embodiments wherein R 2 is hydrogen and n is 2.

[0016] The sixth embodiment includes a compound of any one of the first to fifth embodiments wherein the counter ion for R 1 is selected from chloride, mesylate, bicarbonate, fluoride, nitrate, bromide, sulfate, citrate, benzoate, saccharin anion and acetate.

[0017] The seventh embodiment includes a compound of any one of the first to sixth embodiments wherein R 3 is alkoxy.

[0018] The eighth embodiment is R 4It contains a compound of any one of the first to seventh embodiments which is a triphenylphosphonium cation or a quinone-derived ammonium cation such as, for example, [10-(4,5-dimethyl-3,6-dioxocyclohexane-1,4-dien-1-yl)decyl](tributyl)ammonium bromide.

[0019] The ninth embodiment contains a compound of any one of the first to eighth embodiments in which the counter ion to R 4 is tetrafluoroborate.

[0020] The tenth embodiment contains a compound of any one of the first to ninth embodiments, and the compound contains the following compounds:

[0021]

Chemical formula

[0022] The above R is hydrogen, a hydroxy group, or hydrogen, a hydroxy group, C1-C6 alkyl, O, N, or S which may be substituted with C1-C6 alkyl, and / or the above compound

[0023]

Chemical formula

[0024]

Chemical formula

[0025] The eleventh embodiment contains a method for synthesizing a compound of any one of the first to tenth embodiments.

[0026] The twelfth embodiment contains a composition containing a compound of any one of the first to tenth embodiments and a pharmaceutically acceptable carrier.

[0027] A thirteenth embodiment includes administering the composition of the twelfth embodiment to a human or animal to alleviate conditions caused by inflammation and / or oxidative stress. [Brief explanation of the drawings]

[0028] [Figure 1] FIG. 1 is a graph showing the change in mean plasma concentration (ng / mL) of 2-HOBA over time. [Figure 2] Figure 2. Graph showing the change in mean plasma concentrations of salicylic acid after six ascending single oral doses of 2-hydroxybenzylamine acetate in healthy subjects (n=3 per dose level). See also Table 4. [Figure 3] Figure 3. Schematic for synthesizing TPP linked to the amine group of salicylamine. DETAILED DESCRIPTION OF THE INVENTION

[0029] For the purposes of promoting an understanding of the principles of the novel technology, reference will now be made to preferred embodiments and specific language will be used to describe them. Nonetheless, no limitation on the scope of the novel technology is intended thereby, and it will be understood that such changes, modifications, and further applications as normally occur with respect to the novel technology are within the scope of the disclosure and claims.

[0030] As used herein, unless expressly stated or clearly meant otherwise, the term "about" refers to a range of values of plus or minus 10 percent, e.g., about 1.0 includes values from 0.9 to 1.1.

[0031] As used herein, unless explicitly stated otherwise or clearly having a different meaning, terms such as "therapeutically effective dose" and "therapeutically effective amount" refer to a portion of a compound that has a net positive effect on the health and well-being of humans or other animals. Therapeutic effects can include improvements such as longevity, quality of life, etc., and these effects can also include a decrease in susceptibility to the onset of disease or deterioration of health and well-being. A therapeutic effect can be realized immediately after a single administration and / or treatment, or can be realized cumulatively after a series of administrations and / or treatments.

[0032] Pharmaceutically acceptable salts include salts of the compounds of the present disclosure that are safe and effective for use in mammals and have the desired therapeutic activity. Pharmaceutically acceptable salts include salts of acidic or basic groups present in the compounds of the present disclosure. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Certain compounds of the present disclosure can form pharmaceutically acceptable salts with various amino acids. Suitable base salts include, but are not limited to, aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, and diethanolamine salts. For additional information regarding some pharmaceutically acceptable salts that can be used to practice the present disclosure, see Berge, et al., 66 J.PHARM. SCI. 1-19 (1977), Haynes, et al, J.Pharma. Sci., Vol. 94, No. 10, Oct. 2005, pgs. 2111-2120, etc.

[0033] Both inflammation and oxidative stress can generate bicyclic endoperoxides (prostaglandin H2 and H2-isoprostane, respectively) that can rearrange non-enzymatically to form highly reactive γ-ketoaldehydes (γKAs) in other species. These γKAs, called isolevuglandins and isoketals, covalently modify cellular proteins, for example, via addition to lysine residues, and covalently modify phosphatidylethanolamine by binding to free amines. The levels of γKA protein adducts are thought to increase in many conditions associated with inflammation and oxidative stress, including Alzheimer's disease, atherosclerosis, myocardial infarction, end-stage renal disease, sepsis, atrial fibrillation, chronic kidney disease, radiation-induced tissue damage, and hyperoxia. The development of selective inhibitors that inhibit the endogenous formation of γKA protein adducts in vivo would provide important benefits as therapeutic options for various conditions associated with oxidative stress and inflammation.

[0034] Lipophilic analogs of pyridoxamine (PM) such as salicylamine (SA) and derivatives of SA are highly suitable for this problem. By way of illustration and not limitation, these molecules act as γKA scavengers, inhibiting protein modification and being able to react preferentially with γKA over other lipid carbonyls generated by the peroxidation of hydroxy nonenal. In vitro, the reaction rate of PM and its analogs with γKA is more than 1000 times that of the reaction rate of lysyl residues. However, since lipid peroxidation forms γKA esterified to phospholipids in situ, lipophilic analogs are more effective than the highly hydrophilic PM. Along these lines, SA, a lipophilic PM analog, is highly superior to PM as an inhibitor of γKA protein adducts intracellularly. Furthermore, SA protects HepG2 cells from the cytotoxicity induced by hydrogen peroxide, while PM has no effect. Similarly, SA protects the γKA and oxidation-induced sodium channel function inhibited by γKA or oxidizing agents. The protective effect of SA as an inhibitor of γKA-induced protein modification in cultured cells provides clear evidence of its biological importance in vivo. These results further suggest that inhibiting γKA using SA and SA derivatives is a useful therapeutic strategy for the treatment of diseases.

[0035] In eukaryotic cells, mitochondria are the site of oxidative phosphorylation, and in this capacity, mitochondria are at risk of both suffering from oxidative damage and generating oxidative damage. Targeted delivery of SA and SA derivatives, which exhibit antioxidant properties, to mitochondria allows the molecules to be administered at low doses, thus avoiding side effects due to achieving therapeutic results using high doses while obtaining desirable therapeutic outcomes.

[0036] SA has a very short half-life in the blood, estimated to be about 120 minutes. Due to the short half-life in the bloodstream, SA needs to be administered at least three times to maintain the therapeutic level of SA in the bloodstream. Furthermore, salicylic amine in the blood appears to be processed into salicylic acid. Salicylic acid in the blood has a longer half-life than salicylic amine. Therefore, repeated administration of SA over time can result in the accumulation of very high levels of salicylic acid. Additionally, high levels of salicylic acid can change the metabolism of SA and reduce the therapeutic effect of SA.

[0037] By way of illustration and not limitation, when salicylic amine acts on mitochondria, better therapeutic results may be obtained by directing a higher proportion of the SA administered to the patient towards the mitochondria. By directly targeting the mitochondria via the TPP-SA conjugate disclosed herein, the time SA is exposed to the blood is shortened, a higher effective concentration of SA in the mitochondria is generated, and it becomes possible to deliver a lower dose of the SA agent.

[0038] The accumulation of salicylic acid in the blood can also cause toxicity and adverse effects at very high levels. The half-life of salicylic acid in the blood is 2 - 12 hours depending on the amount used or produced. Approximately 80 - 90% of all salicylates in the blood are protein-bound, and the rest are in the free form, which is detectable by normal analysis. Most of the excretion of circulating salicylates occurs via urine. Since a blood concentration of 35 mg / dL or higher is considered toxic, there is a sufficient possibility of death if salicylic acid accumulates further in the body. In many cases, toxicity appears within a few days after use. The most severe cases leading to coma and death occur in psoriasis patients who have applied a very large amount of salicylic acid topically to the skin. If the composition can be directly targeted to the mitochondria, the accumulation of salicylic acid will not occur and the dosage can be made lower.

[0039] Put simply, the currently taught safe dosage of SA in the art results in an increase in the plasma level of SA. When SA is present at high levels in the bloodstream, a portion of the SA derived from the blood can enter the cells, and ultimately, a portion of the SA administered to the patient can enter the mitochondria.

[0040] Some strategies for preferentially or selectively delivering molecules to mitochondria utilize the substantial negative electrochemical potential maintained across the inner mitochondrial membrane. Delocalized lipophilic cations are particularly effective at crossing hydrophobic membranes and thus preferentially accumulate within the mitochondrial matrix. Some molecules that utilize this approach contain a headpiece of a triphenylphosphonium (TPP) salt attached to the molecule to be delivered to the mitochondria. TPP has the ability to be transported into the mitochondria against a concentration gradient. In other words, by binding a compound to TPP, a much higher concentration is achieved within the mitochondria than in the cytoplasm or blood. In some aspects of the present disclosure described herein, the dosage of TPP-conjugated SA required is much less than that of SA alone and thus reduces potential toxicity or other undesirable side effects.

[0041] In some aspects of the present disclosure disclosed herein, the salicylamine derivatives target mitochondria directly. This class of molecules does not rely on antioxidant action to suppress the formation of γKA but rather acts as a surrogate amine for the adduct to prevent the formation of γKA protein adducts. This new group of compounds provides both a valuable tool for further investigating biological pathways and a potentially powerful treatment for conditions caused by inflammation and oxidative stress.

[0042] In some embodiments, TPP is conjugated to a salicylamine derivative to target the delivery of salicylamine to the mitochondria of cells. The TPP moiety may be conjugated at different positions of the salicylamine ring and may be conjugated via different atoms. In one embodiment of the present disclosure, (5-((2-hydroxylbenzyl)amino)pentyl)triphenylphosphonium bromide is synthesized as shown in Scheme 1 below.

[0043] Scheme 1: Synthesis of (5-((2-hydroxylbenzyl)amino)pentyl)triphenylphosphonium bromide

[0044]

Chem.

[0045] Referring now to Figure 3, starting from 5-amino-1-pentanol, the desired triphenylphosphonium salt is synthesized in three steps. The alcohol is treated with concentrated hydrobromic acid to provide the desired halogen in quantitative yield. The primary halide reacts with triphenylphosphine (PPh3) in acetonitrile (MeCN) to produce the triphenylphosphonium salt in quantitative yield. Reduction amination with the triphenylphosphonium salt and salicylaldehyde in methanol (MeOH) follows to form the desired triphenylphosphonium salt.

[0046] In a further embodiment of the invention, the cation can be conjugated to both via different positions on the ring and different atoms. For example, a dicationic species in which the triphenylphosphonium cation is conjugated to the para oxygen atom with respect to the hydroxyl group is synthesized as shown in Scheme 2 below.

[0047] Scheme 2: Synthesis of (5-(allyloxy)-2-hydroxyphenyl)methanaminium acetate

[0048]

Chem.

[0049] The hydroxyl group at the 5-position is selectively allylated using allyl bromide and sodium carbonate. Treatment with hydroxylamine hydrochloride and sodium hydroxide in water gives the corresponding oxime, which is reduced to the amine with zinc in acetic acid.

[0050] Scheme 3: Synthesis of (3-(3-(ammoniomethyl)-4-hydroxyphenoxy)propyl)triphenylphosphonium tetrafluoroborate

[0051] [ka]

[0052] The salt is treated with triphenylphosphonium tetrafluoroborate in the presence of triphenylphosphine, 1,1'-azobis(cyclohexanecarbonitrile) (ACN), and acetic acid to give the desired dicationic species.

[0053] Scheme 4: Synthesis of deuterated salts

[0054] [ka]

[0055] Starting from hydroquinone, tetradeuteration can be achieved using deuterated acid and deuterated methanol in deuterium oxide. From there, formylation can be carried out using triethylamine (EtN), magnesium chloride, and paraformaldehyde in acetonitrile. Using deuterated 2,5-dihydroxybenzaldehyde XIII, the desired deuterated salt can be synthesized using the synthetic methods described above.

[0056] In animal models in which salicylamine in drinking water can be administered to mice, dosages between 1 and 10 grams per liter of water produce tissue concentrations of 10 - 500 micromoles, which fall within the range known to inhibit γKA intracellularly, but high dosages of 1 gram per milliliter of water have been used.

[0057] Other modes of administration of salicylamine have been shown to be effective. Mice administered 200 mg / kg of salicylamine intraperitoneally show a reduction in the edema of the paw injected with carrageenan. Derivatives of salicylamine such as 5-methylsalicylamine are also effective in the management of inflammation. Scheme 5: Synthesis of C-6 derivatives of 2-aminomethylphenol

[0058]

Chemical formula

[0059] R is hydrogen, hydroxy, C1 - C6 alkyl, O, N or S, and may be substituted with hydrogen, hydroxy, or C1 - C6 alkyl.

[0060] These derivatives of 2-aminomethylphenol can be prepared by orthoacylation utilizing the aldol chemistry of phenolate alkoxides. Alternatively, allyl ethers of phenol can be prepared and the Claisen rearrangement can be used to insert an allyl group at the 6-position. These new compounds contain functionalities that allow the preparation of conjugates while retaining the aminophenol moiety. Yet another approach is orthocarboxylation via alkoxides, and carbon dioxide is another approach. These compounds contain functionalities that allow the preparation of conjugates while retaining the aminophenol moiety. The compounds depicted in Scheme 5 can be conjugated with a triarylphosphine (phosphonium salt) that facilitates reagents localized in mitochondria. For example, the 6-hydroxymethyl analog can be coupled with a hydroxyalkylphosphonium salt using an acid catalyst.

[0061] The term "alkyl" group includes straight-chain or branched saturated aliphatic hydrocarbon chains such as, for example, methyl, ethyl, isopropyl, tert-butyl, etc. The term "alkoxy" group includes straight-chain, branched, or cyclic hydrocarbons bonded to an oxygen atom such as, for example, methoxy, ethoxy, isopropoxy, tert-butoxy, etc.

[0062] The compounds of the present disclosure can be administered as a single active agent or can be used in combination with one or more other agents useful for the treatment or prevention of various complications such as, for example, Alzheimer's disease and other neurodegenerative diseases, hypertension, fatty liver disease, alcohol-related liver disease, chronic obstructive pulmonary disease, pulmonary hypertension, radiation-induced tissue damage, and gastroesophageal reflux disease. The compounds of the present disclosure can be used as a single active agent or in combination with one or more other agents, for example, for the prevention of ischemia-reperfusion injury and the prevention of arrhythmia. When administered in combination, the therapeutic agents can be formulated as separate compositions administered simultaneously or at different times, or the therapeutic agents can be administered as a single composition.

[0063] The compounds of the present disclosure can be constituted in solid form (including granules, powders, or suppositories) or in liquid form (for example, solutions, suspensions, or emulsions). They can be applied to various solutions, subjected to conventional pharmaceutical operations such as sterilization, and / or can contain conventional adjuvants such as preservatives, stabilizers, wetting agents, emulsifying agents, buffer solutions, etc.

[0064] For administration, the compounds of the present disclosure are usually combined with one or more adjuvants. For example, they can be mixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acids, stearic acid, talc, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric acid and sulfuric acid, acacia, gelatin, sodium alginate, polyvinylpyrrolidine, and / or polyvinyl alcohol and can be tableted or encapsulated for conventional administration.

[0065] Alternatively, they may be dissolved in physiological saline, water, polyethylene glycol, propylene glycol, carboxymethyl cellulose colloidal solution, ethanol, corn oil, peanut oil, cottonseed oil, sesame oil, tragacanth gum and / or various buffer solutions. Other adjuvants and modes of administration are well known in the pharmaceutical art. The carrier or diluent may include a time-delay material such as glyceryl monostearate or glyceryl distearate, alone or with waxes, or other materials well known in the art.

[0066] In therapeutic use, the compounds of the present disclosure can be administered to mammalian patients in an amount sufficient to reduce or inhibit the desired symptoms. The effective amount for this use depends on factors including, but not limited to, the route of administration, the stage and severity of the indication, the general health of the mammal, and the judgment of the prescribing physician. The compounds of the present disclosure are safe and effective over a wide range of dosages. However, it is understood that the actual amount of pyridoxamine administered will be determined by the physician in light of the above relevant circumstances.

[0067] The compounds described herein can be formulated into dosage unit forms containing vehicles including conventional pharmaceutically acceptable carriers, adjuvants, and liposomes, and can be administered by any suitable route including oral, enteral, parentally, inhalation, or rectal. As used herein, the term parenteral includes subcutaneous, intravenous, intraarterial, intramuscular, intrasternal, intratendinous, intraspinal, intracranial, intrathoracic, infusion techniques, within dental caries, enteral, or intraperitoneal.

Examples

[0068] Example Example 1: Synthesis of (5-((2-Hydroxybenzyl)amino)pentyl)triphenylphosphonium Bromide (V)

[0069]

Chemical formula

[0070] Reaction of 5-amino-1-pentanol I with concentrated hydrobromic acid produced the desired halogen II in quantitative yield. The bromide II was then reacted with triphenylphosphine in acetonitrile to produce the triphenylphosphonium salt III in quantitative yield. Reductive amination of the triphenylphosphonium salt III with salicylaldehyde IV in the presence of sodium borohydride in methanol afforded the triphenylphosphonium salt V in 45% yield based on recovered starting material.

[0071] Referring now to Figure 3, further details regarding the synthesis of TPP attached to the amine group of salicylamine are provided.

[0072] Example 2: Synthesis of (5-(allyloxy)-2-hydroxyphenol)methanaminium acetate (IX):

[0073] [ka]

[0074] To a 25 mL round-bottom flask was added 2,5-dihydroxybenzaldehyde VI (1.48 g, 10.7 mmol), acetonitrile (MeCN) (11 mL), sodium carbonate (2.27 g, 21.4 mmol), and allyl bromide (0.93 mL, 10.7 mmol). The reaction was heated to 80 °C and refluxed overnight. The resulting solution was cooled to room temperature, poured into 20 mL of 1 M NaOH, and extracted with 20 mL of ethyl acetate (EtOAc) to remove the 2,5-bis(allyloxy)benzaldehyde by-product. The aqueous layer was acidified to pH 1 with concentrated HCl and extracted 3 × 20 mL with EtOAc. The organic layer was dried over MgSO and filtered. The solvent was removed under reduced pressure. The product was purified by column chromatography using ethyl acetate:hexane=1:4 to give 5-(allyloxy)-2-hydroxybenzaldehyde VII (0.35 g, 1.99 mmol) in 19% yield.

[0075] To a 25 mL round-bottom flask, 5-(allyloxy)-2-hydroxybenzaldehyde VII (0.16 g, 0.9 mmol) was added. Hydroxylamine hydrochloride (0.10 g, 1.37 mmol) and sodium hydroxide (0.06 g, 1.37 mmol) were dissolved in 1.5 mL of deionized water. This aqueous solution was added, and the reaction mixture was heated at 80 °C for 1 hour. The mixture was cooled to room temperature and poured into 20 mL of HCl solution (pH 1). The solution was extracted with 3 × 20 mL of EtOAc, dried over MgSO4, and filtered. The solvent was removed under reduced pressure to obtain 5-(allyloxy)-2-hydroxybenzaldehyde oxime VIII (0.17 g, 0.9 mmol) in quantitative yield.

[0076] To a 10 mL round-bottom flask, 5-(allyloxy)-2-hydroxybenzaldehyde oxime VIII (0.17 g, 0.9 mmol) was added, followed by acetic acid (AcOH) (2 mL) and zinc powder (0.20 g, 3 mmol). The reaction mixture was stirred at room temperature overnight. The solution was diluted with methanol (5 mL), the zinc powder was filtered off, and the solvent was removed under reduced pressure. To remove traces of acetic acid, it was necessary to wash several times with toluene and then remove the solvent under reduced pressure. (5-(allyloxy)-2-hydroxyphenyl)methanaminium acetate IX (0.21 g, 0.9 mmol) was recovered in quantitative yield.

[0077] Example 3: Synthesis of Triphenylphosphonium Tetrafluoroborate:

[0078] To a 125 mL Erlenmeyer flask, triphenylphosphine (PPh3) (2.91 g, 11 mmol) was added and dissolved in diethyl ether (Et2O) (15 mL). When tetrafluoroborate diethyl ether complex (1.36 mL, 10 mmol) was added, a white precipitate was formed. The precipitate was collected by filtration and recrystallized from chloroform to obtain triphenylphosphonium tetrafluoroborate (0.97 g, 2.7 mmol) in 27% yield.

[0079] Example 4: Synthesis of (3-(3-(Ammoniomethyl)-4-hydroxyphenoxy)propyl)triphenylphosphonium acetate tetrafluoroborate (X):

[0080] [Chemical formula]

[0081] To a 50 mL round-bottom flask were added (5-(allyloxy)-2-hydroxyphenol)methanaminium acetate IX (0.14 g, 0.6 mmol), chlorobenzene (PhCl) (25 mL), and acetic acid (2 mL). After the compound was completely dissolved, 1,1'-azobis(cyclohexanecarbonitrile) (0.03 g, 0.12 mmol), triphenylphosphonium tetrafluoroborate (0.51 g, 2.64 mmol), and triphenylphosphine (0.03 g, 0.12 mmol) were added. The reaction vessel was sealed with a septum and purged with argon gas for 5 minutes. The reaction mixture was heated to 110 °C under balloon pressure and reacted overnight. The mixture was cooled to room temperature and the solvent was removed under reduced pressure. It was necessary to wash several times with toluene to remove a small amount of acetic acid and chlorobenzene, and then remove the solvent under reduced pressure. The obtained crude solid was triturated several times with chloroform to obtain (3-(3-(ammoniomethyl)-4-hydroxyphenoxy)propyl)triphenylphosphonium acetate tetrafluoroborate X (0.14 g, 0.29 mmol) in a 48% yield.

[0082] Example 5: Synthesis of Triphenylphosphonium (TPP) Bonded to the Amino Group of Salicylamide

[0083] [Chemical formula]

[0084] To a 25 mL round-bottom flask, 1.11 g of 5-amino-1-pentanol (10 mmol) was added. 10 mL of 48% hydrogen bromide in water was added. The reaction vessel was refluxed for 3 hours. Removal of the solvent under reduced pressure gave a brown, viscous solid. This compound was used in the next step without further purification. To a 100 mL round-bottom flask, 2.47 g of 5-bromopentane-1-amine hydrobromide (10 mmol) was placed. 50 mL of acetonitrile was added to the flask. 5.27 g of triphenylphosphine (20 mmol) was added to the flask. The flask was heated under reflux for 60 hours. Removal of the solvent under reduced pressure gave a brown crude oil. The oil was dissolved in 30 mL of water and washed with 3 × 30 mL of diethyl ether. The aqueous phase was basified with sodium carbonate and then extracted with 3 × 30 mL of dichloromethane. The solvent was removed under reduced pressure. To a 10 mL round-bottom flask, 0.30 g of (5-aminopentyl)triphenylphosphonium bromide (0.7 mmol) was placed. 5 mL of methanol was added. 0.07 mL of salicaldehyde (0.7 mmol) was added. The flask was stirred overnight. 0.04 g of sodium borohydride (1.05 mmol, 1.5 eq) was added to the flask. The reaction was allowed to proceed until gas evolution ceased. The solution was poured into 20 mL of water and extracted with 3 × 20 mL of dichloromethane. The solvent was removed under reduced pressure. The crude solid was dissolved in the minimum amount of dichloromethane and diethyl ether was allowed to overflow. White crystals collected to form a mixture of products in a 4:5 ratio.

[0085] Example 6: Dosage Guidance Study

[0086] Subjects Healthy male and non-pregnant female volunteers over 18 years of age were eligible to participate. Subjects were not permitted to take medications two weeks prior to or during the study. Exclusion criteria included Known heart, kidney, or liver disease; The presence of a disease with a definite medical condition or symptoms / signs that could confound the interpretation of the study results; The need to discontinue medications administered as standard treatment; and The desire or inability to use an approved contraceptive method was included.

[0087] Compound 2-Hydroxybenzylamine (as acetate, CAS 1206675-01-5) was obtained from TSI (China) Co., Ltd. (Shanghai, China). A commercial production lot was used (Lot 16120312). In our laboratory, HPLC and NMR spectroscopy were used to confirm that the purity of the commercial lot exceeded 99%. Hard gel capsules (Capsugel, Jiangsu, China) containing 50, 110, and 412.5 mg of 2-hydroxybenzylamine acetate (equivalent to 34, 75, and 281 mg of 2-hydroxybenzylamine) were prepared by TSI (China) Co., Ltd. The measured values of the average fill weight, weight uniformity, disintegration, 2-hydroxybenzylamine content, acetate content, and microbiological and analytical tests were all within the specification ranges.

[0088] Study design This study was an open-label, single ascending dose study designed to evaluate the pharmacokinetics, safety, and tolerability of a single dose of 2-hydroxybenzylamine acetate. A 3 + 3 clinical trial design using a modified Fibonacci dosing scheme 14 was used, with a starting dose of 50 mg. Subsequently, the doses were increased to 100, 200, 330, 550, and 825 mg. These doses of 2-hydroxybenzylamine acetate correspond to 34, 68, 136, 224, 373, and 560 mg of 2-hydroxybenzylamine. Each increase in dose was initiated only after reviewing the safety data from all subjects who had received the previous dose.

[0089] The subjects were admitted to the Vanderbilt University Clinical Research Center and stayed in the 24-hour unit after being orally administered 2-hydroxybenzylamine acetate in capsules. This study did not include a placebo control, and neither the staff nurses nor the subjects were informed about the content of the capsule administration. The subjects were monitored at intervals defined in the protocol for 24 hours after the administration of 2-hydroxybenzylamine. Safety evaluations included vital signs (heart rate, respiratory rate, blood pressure, and SpO2), clinical laboratory parameters (blood biochemistry, hematology, and urine tests), 12-lead electrocardiogram (ECG), and assessment of potential adverse events. All adverse events were recorded regardless of whether they were considered related to the study.

[0090] Pharmacokinetic Sampling and Analysis Blood samples for pharmacokinetic analysis were collected at baseline, 0.25, 0.5, 1, 2, 6, 4, 8, and 24 hours after the administration of 2-hydroxybenzylamine acetate at all doses. Samples at 0.25 hours were collected only at doses of 200 mg or less, and samples at 6 hours were collected only at doses of 330 mg or more. At each time point, the plasma concentrations of 2-hydroxybenzylamine and salicylic acid, the primary metabolite of 2-hydroxybenzylamine, were measured.

[0091] As previously described, 2 H4]-2-hydroxybenzylamine prepared by Dr. Venkataraman Amarnath was used as the internal standard. 2An internal standard solution of [H4]-2-hydroxybenzylamine (100 ng / mL) was prepared in acetonitrile and added to all standard samples, quality control samples, and patient samples. Standard samples and quality control samples of 2-hydroxybenzylamine at 1 mg / mL were prepared in water. Eight standard curve samples (5, 10, 20, 100, 200, 1000, 2000, and 5000 ng / mL) were prepared using blank human plasma (Bioreclamation, Westbury, NY). Additionally, three quality control samples (15, 300, and 3000 ng / mL) were prepared in blank human plasma. Plasma samples were thawed at room temperature and vortexed. An internal standard solution (400 μL) was added to either 100 μL of plasma, quality control, or standard sample and mixed in a protein precipitation filter 96-well plate (Phenomenex, Torrance, CA). This solution was eluted into a 96-well plate using a positive pressure manifold and dried at 40 °C under nitrogen gas. Next, the samples were reconstituted with 97 / 3 v / v water / acetonitrile containing 10 mM ammonium formate for analysis. Liquid chromatography tandem mass spectrometry of 2-hydroxybenzylamine was performed using a Shimadzu Nexera X2 LC-30AD pump, column oven, and degasser (Kyoto, Japan) (column: C18 2.1×50 mm, 1.7 μm, Phenomenex, Torrance, CA) combined with a Sciex QTrap 5500 mass spectrometer equipped with a TurboV ion source (Framingham, MA). Quantification of 2-hydroxybenzylamine was performed using electrospray ionization in positive ionization mode. The column temperature was set at 60 °C and the flow rate was 0.5 mL / min. An aqueous solution of 10 mM ammonium formate was used as mobile phase A and a 1% formic acid-acetonitrile solution was used as mobile phase B, and a gradient of 3-90% B (A 97-10%) from 0 to 0.90 minutes was set. Quantification of 2-hydroxybenzylamine was verified in the range of 5-5000 ng / mL, with intra-assay precision of 3.7-7.0%, bias of -9.7-2.8, inter-assay precision of 4.4-6.2%, and bias of -7.1-1.64.All standard samples and quality control samples met the acceptance criteria (standard curve R. 2 > 0.90, 66.7% of all QC samples and at least 50% at each concentration within 15% of the nominal concentration).

[0092] Plasma concentration-time data were imported into Phoenix WinNonlin® 8.0 software (Certara USA, Inc., Princeton, NJ), and oral pharmacokinetic parameters of 2-hydroxybenzylamine were estimated from each dose for individual subjects. Non-compartmental analysis using Model 200 (plasma, single extravascular dose, linear log trapezoidal method) was performed for each plasma concentration-time profile, and individual pharmacokinetic parameters (half-life, area under the concentration-time curve (AUC), maximum observed plasma concentration (C max ) and time to reach the maximum observed plasma concentration (T max )) were estimated.

[0093] Statistical Analysis Descriptive statistics (mean, standard deviation, standard error) were used for the evaluation of demographics, safety, and pharmacokinetics. Study Population

[0094]

Table 1

[0095] Refer to Table 1 here. A total of 18 volunteers were enrolled and completed the study successfully (3 subjects at each dose level). The demographics of the subjects are shown in Table 1 and were similar among the dosing groups. Safety

[0096]

Table 2

[0097] Refer to Table 2 here. All reported adverse events are summarized in Table 2. Five participants (28%) reported at least one adverse event during the study. The most frequently reported adverse event (two cases) was frequent urination (two subjects, 11%). All adverse events were mild. There were no adverse events judged to be related to the study, nor was there a dose-dependent increase in the frequency or severity of adverse events. No clinically significant changes related to 2-hydroxybenzylamine were observed in ECG recordings, vital signs, or laboratory parameters. There were no serious adverse events or deaths. Pharmacokinetics

[0098] [Table 3]

[0099] Refer to Table 3 and Figure 1 here. The mean plasma concentration-time profile of 2-hydroxybenzylamine and the estimated pharmacokinetic parameter values are shown in Figure 1 and Table 3, respectively. After a single oral dose of 2-hydroxybenzylamine, dose-dependent changes in the maximum plasma concentration (C max ) and the area under the concentration-time curve (AUC) were observed. The mean time to reach C max was 1.6 hours, and the mean half-life of 2-hydroxybenzylamine was 2.1 hours.

[0100] [Table 4]

[0101] Refer to Table 4 and Figure 2 here. The plasma concentration of salicylic acid, a primary metabolite of 2.2-hydroxybenzylamine, was also measured. The systemic exposure to salicylic acid after a single oral dose of 2-hydroxybenzylamine acetate at each dose level is shown in Figure 2 and quantified in Table 4. After oral administration of 2-hydroxybenzylamine, dose-dependent changes in the systemic exposure (C max and AUC) of salicylic acid were observed. The T maxwas in the range of 2.67 to 4.67 hours and tended to increase as the dosage of 2-hydroxybenzylamine increased.

[0102] Literature 1. Amarnath, V.; Amarnath, K.; Amarnath, K.; Davies, S.; Roberts, L.J., 2nd. Pyridoxamine: an extremely potent scavenger of 1,4-dicarbonyls. Chem. Res. Toxicol. 2004, 17, 410 - 415. 2. Brame, C.J.; Salomon, R.G.; Morrow, J.D.; Roberts, L.J., 2nd. Identification of extremely reactive gamma-ketoaldehydes (isolevuglandins) as products of the isoprostane pathway and characterization of their lysyl protein adducts. J. Biol. Chem. 1999, 274, 13139 - 13146. 3. Brame, C.J.; Boutaud, O.; Davies, S.S.; Yang, T.; Oates, J.A.; Roden, D.; Roberts, L.J., 2nd. Modification of proteins by isoketal-containing oxidized phospholipids. J. Biol. Chem. 2004, 279, 13447 - 13451. 4. Davies, S.S.; Talati, M.; Wang, X.; Mernaugh, R.L.; Amarnath, V.; Fessel, J.; Meyrick, B.O.; Sheller, J.; Roberts, L.J., 2nd. Localization of isoketal adducts in vivousing a single-chain antibody. Free Radic. Biol. Med. 2004, 36, 1163-1174. 5. Davies, S.S.; Brantley, E.J.; Voziyan, P.A.; Amarnath, V.; Zagol-Ikapitte, I.; Boutaud, O.; Hudson, B.G.; Oates, J.A.; Ii, L.J. Pyridoxamine Analogues Scavenge Lipid-Derived gamma-Ketoaldehydes and Protect against H(2)O(2)-Mediated Cytotoxicity. Biochemistry 2006, 45, 15756-15767. 6. Frantz, M.-C. and Wipf, P. Mitochondria as a target in treatment. Environ. Mol. Mutagen 2010, 51, 462-475. 7.Fukuda, K.; Davies, S.S.; Nakajima, T.; Ong, B.H.; Kupershmidt, S.; Fessel, J.; Amarnath, V.; Anderson, M.E.; Boyden, P.A.; Viswanathan, P.C.; Roberts, L.J., 2nd; Balser, J.R. Oxidative mediated lipid peroxidation recapitulates proarrhythmic effects on cardiac sodium channels. Circ. Res. 2005, 97, 1262-1269. 8.Hoppe, G.; Subbanagounder, G.; O'Neil, J.; Salomon, R.G.; Hoff, H.F. Macrophage recognition of LDL modified by levuglandin E2, an oxidation product of arachidonic acid. Biochim. Biophys. Acta 1997, 1344, 1-5. 9.Madan, R.; Levitt, J. A review of toxicity from topical salicylic acid preparations. J. Amer. Acad. Dermatol. 2014, 70, 788-92. 10.Nakajima, T.; Davies, S.S.; Matafonova, E.; Potet, F.; Amarnath, V.; Tallman, K.A.; Serwa, R.A.; Porter, N.A.; Balser, J.R.; Kupershmidt, S.; Roberts, L.J., II. Selective gamma-ketoaldehyde scavengers protect NaV1.5 from oxidant-induced inactivation. J. Mol. Cell. Cardiol. 2010, 48, 352-359. 11.Poliakov, E.; Brennan, M.L.; Macpherson, J.; Zhang, R.; Sha, W.; Narine, L.; Salomon, R.G.; Hazen, S.L. Isolevuglandins, a novel class of isoprostenoid derivatives, function as integrated sensors of oxidant stress and are generated by myeloperoxidase in vivo. Faseb J 2003, 17, 2209-2220. 12.Salomon, R.G.; Miller, D.B.; Zagorski, M.G.; Coughlin, D.J. Solvent Induced Fragmentation of Prostaglandin Endoperoxides. New Aldehyde Products from PGH2 and Novel Intramolecular 1,2-Hydride Shift During Endoperoxide Fragmentation in Aqueous Solution. J. Am. Chem. Soc. 1984, 106, 6049-6060. 13.Salomon, R.G.; Batyreva, E.; Kaur, K.; Sprecher, D.L.; Schreiber, M.J.; Crabb, J.W.; Penn, M.S.; DiCorletoe, A.M.; Hazen, S.L.; Podrez, E.A. Isolevuglandin-protein adducts in humans: products of free radical-induced lipid oxidation through the isoprostane pathway. Biochim. Biophys. Acta 2000, 1485, 225-235. 14.Sullivan, C.B.; Matafonova, E.; Roberts, L.J., II; Amarnath, V.; Davies, S.S. Isoketals form cytotoxic phosphatidylethanolamine adducts in cells. J. Lipid Res. 2010,51, 999-1009. 15.Zagol-Ikapitte, I.; Masterson, T.S.; Amarnath, V.; Montine, T.J.; Andreasson, K.I.; Boutaud, O.; Oates, J. A. Prostaglandin H(2)-derived adducts of proteins correlate with Alzheimer's disease severity. J. Neurochem. 2005, 94, 1140-1145. 16.Zagol-Ikapitte, I., et al. Determination of the pharmacokinetics and oral bioavailability of salicylamine, a potent gamma-ketoaldehyde scavenger, by LC / MS / MS. Pharmaceutics 2010, 2, 18-29.

[0103] The novel technology has been described and set forth in detail in the foregoing description, which is to be regarded as illustrative only and not as limiting in nature, and it is understood that only the preferred embodiments have been shown and described. And it is understood that all changes and modifications within the scope of the spirit of the novel technology are desired to be protected. Similarly, the new technology has been described using specific examples, theoretical discussions, evaluations, and diagrams, but these diagrams and the accompanying discussions should in no way be construed as limiting the technology. All patents, patent applications, and references to texts, scientific papers, publications, etc. referred to in this application are hereby incorporated by reference in their entirety into this specification.

Claims

1. The following formula (I): 【Chemical 1】 (wherein R 1 may be hydrogen which may have a counter ion, R 2 is an acyl group or an alkyl group substituted with R 4 , R 3 is selected from hydrogen; halogen; hydroxy group; acyl group; alkoxy group; an alkyl group which may be substituted with an alkyl group, alkenyl group, hydroxy group, acyl group or alkoxy group, a C 1 -C 6 alkyl group; a carbonyl group which may be substituted with hydrogen, hydroxy group, acyl group or a C 1 -C 3 alkyl group, R 3 may or may not be substituted, and when substituted, the substituent is hydrogen, hydroxy group or a C 4 -C 1 -C 3 alkyl group which may be substituted with each R 4 is a triphenylphosphonium cation having a counter ion, and n is 1 or 2).

2. Said R 3 is hydrogen, and said R 2 is an alkyl group substituted with R 4 The compound according to claim 1.

3. Said R 2 is R 4 substituted C 5 is an alkyl group of the compound according to claim 2.

4. The compound according to claim 3, wherein the counter ion is bromide.

5. Said R 2 The compound according to claim 1, wherein R is hydrogen and n is 2.

6. The counter ion to said R 1 is selected from chloride, mesylate, bicarbonate, fluoride, nitrate, bromide, sulfate, citrate, benzoate, saccharin anion, and acetate, and the compound according to claim 5.

7. The aforementioned R 1 The compound according to claim 6, wherein the counter ion to R is an acetate.

8. Said R 3 The compound according to claim 5, wherein R is an alkoxy group.

9. The aforementioned R 4 The compound according to claim 7, wherein the counter ion to R is tetrafluoroborate.

10. A composition comprising the compound according to any one of claims 1 to 9 and a pharmaceutically acceptable carrier.

11. A composition for reducing side effects caused by salicylamine, comprising the compound according to any one of claims 1 to 9 and a pharmaceutically acceptable carrier.

12. A composition for reducing inflammation and oxidative stress, comprising the compound according to any one of claims 1 to 9 and a pharmaceutically acceptable carrier.

13. The composition according to any one of claims 10 to 12 for administration to a human or an animal.

14. The following formula (II): 【Chemical 2】 (5-((2-Hydroxybenzyl)amino)pentyl)triphenylphosphonium bromide having the structural formula of

15. The following formula (III): [Chemical Formula 3] (5-((2-Hydroxyphenyl)amino)pentyl)triphenylphosphonium bromide having the structural formula of

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