Preparation and use of stable dephosphorized operin salts, esters and conjugates

Abstract

The present invention provides pharmaceutically acceptable salts, esters, and conjugates of dephosphorized luminocap, compositions, methods for their production, and uses in the treatment of diseases or conditions treatable with dephosphorized luminocap or luminocap.

Description

[0001] This patent application claims the benefit of priority to U.S. Provisional Application Serial No. 63 / 388,414, filed on July 12, 2022, the teachings of which are incorporated herein by reference in their entirety. Technical Field

[0002] Disclosed are dephosphocyanidin salts, esters and conjugates having improved stability, physical properties and / or handling characteristics compared to dephosphocyanidin, pharmaceutical compositions containing these dephosphocyanidin salts, esters and conjugates, and methods of using them in conditions that can be treated with dephosphocyanidin. Background Art

[0003] The structure of the cypridine represented in formula I

[0004]

[0005] is a phosphate prodrug of dephosphocyanidin as depicted in Formula II

[0006] .

[0007] When applied to a subject, chlorophyll is metabolized to form dephosphorylated chlorophyll. Chlorophyll undergoes acid-catalyzed and / or enzymatic dephosphorylation reactions, resulting in the loss of a phosphate group, exposing the hydroxyl group of dephosphorylated chlorophyll. Chlorophyll exists as a zwitterion, in which the phosphoric acid and amine moieties are mutually ionized. The presence of zwitterions limits the solubility of chlorophyll, and also limits its ability to form salts with the optional acid that can exist under physiological tolerance conditions. Removing the phosphate group allows the formation of an intermolecular salt of dephosphorylated chlorophyll, which was originally impossible to prepare with chlorophyll. As a non-ionized form, dephosphorylated chlorophyll is much more lipophilic than chlorophyll, and therefore can more effectively cross the blood-brain barrier to elicit a response. Dephosphorylated chlorophyll has a high affinity to the 5-HT2A receptor, and can activate the 5-HT2A receptor, which plays a key role in regulating emotions, sexual behavior, aggression, impulsivity, cognitive function, appetite, pain, sleep and memory, and other behaviors.

[0008] Evidence of therapeutic efficacy of dephosphocyanidins in a wide range of clinical applications, including psychiatric disorders, pain disorders, and neurological disorders, has generated considerable interest in this compound. However, the unfavorable physical properties of dephosphocyanidins in the solid state, such as poor crystallinity, limited improvement in bulk purity after crystallization, susceptibility to autocatalytic oxidation after manipulation and long-term storage, and low aqueous solubility, have hampered the development of dephosphocyanidin-based drugs.

[0009] Classical psychedelics such as dephosphocybin, which are serotonergic hallucinogens, have been shown in multiple lines of research to potentially induce therapeutic changes in people with various psychiatric disorders. However, since natural dephosphocybin is present only in relatively small amounts in actual mushrooms (Tylš et al., European Neuropsychopharmacology 2014 24(3): 342–356), and unmodified dephosphocybin is relatively unstable in solution, most research has focused on prodrugs of dephosphocybin, which are metabolized to dephosphocybin after ingestion and are responsible for the demonstrated therapeutic and behavioral effects.

[0010] Results of completed and published studies have shown that dephosphocyanin exposure (via cyanocobalamin administration) results in significant improvements in symptoms of anxiety, depression, and substance use disorders.

[0011] In addition, the clinical safety of dephosphocyanidin has been extensively studied in adult populations, both as a single agent and as adjunctive therapy. Dephosphocyanidin is most commonly administered as cyanidin capsules by oral administration and has been evaluated in open-label and double-blind controlled trials. The dosage regimen of cyanidin ranges from 0.014 mg / kg to 0.6 mg / kg (based on an estimated dose-normalized bioavailability of 52.7% from (F. Hasler et al., Pharmaceutica Acta Helvetiae 1997 72(3), 175–184), roughly corresponding to a dose range of 7 µg / kg - 0.32 mg / kg dephosphocyanidin), administered as a single dose or in multiple ascending doses spaced several weeks apart.

[0012] Among the adverse psychological experiences, anxiety, induction of negative emotional states, and paranoid or delusional thinking are frequently reported. In terms of physical adverse events, they are cardiovascular events (i.e., increases in blood pressure and heart rate) as well as nausea and headache. In addition, hallucinogenic effects with increased anxiety have been reported at doses as low as 5 mg of cyprid (equivalent to approximately 2 mg of dephosphocyprid). See, e.g., Griffith et al., Pharmaceutica Acta Helvetiae 2011 72(3) 175–184.

[0013] US Patent No. 11312684 discloses that dephosphorylation cyanidin benzoate and dephosphorylation cyanidin succinate are preferred salt forms for producing pharmaceutical compositions with excellent shelf life stability and resistance to oxidative degradation. Some of the salts are disclosed to have stability of up to three weeks.

[0014] There is a need for dephosphocyanidin salts, esters and conjugates and formulations thereof having improved stability, physical properties and / or handling characteristics compared to dephosphocyanidin. Summary of the invention

[0015] One aspect of the present disclosure relates to the preparation of dephosphorylated cyperus salts, esters and conjugates.

[0016] Another aspect of the present disclosure relates to pharmaceutical compositions comprising these dephosphorylated cypridine salts, esters and conjugates.

[0017] Another aspect of the present disclosure relates to methods of producing these dephosphorylated cypridine salts, esters and conjugates.

[0018] Yet another aspect of the present disclosure relates to methods of using these dephosphocyanidin salts, esters and conjugates and pharmaceutical compositions thereof to treat conditions and diseases treatable with dephosphocyanidin or cyanidin. DETAILED DESCRIPTION

[0019] definition

[0020] To facilitate understanding of the present invention, a number of terms are defined below and throughout this disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those of ordinary skill in the art to which the present invention belongs. The terms herein are used to describe specific embodiments of the present invention, but their usage does not limit the present invention, except as outlined in the claims.

[0021] Unspecific references do not refer to a single entity but include general categories that can be illustrated by specific instances.

[0022] As used herein, the term "about" refers to a value that is greater than or less than within 10% of the stated value.

[0023] The term "administering" refers to a method of providing a dose of a compound or pharmaceutical composition to a subject.

[0024] As used herein, the phrases "pharmacologically effective amount", "therapeutically effective amount", etc., when used to refer to the dephosphorylated cypridine salts, esters and conjugates disclosed herein and pharmaceutical compositions comprising these salts, esters and conjugates, refer to an amount sufficient to alleviate one or more symptoms of the disease or condition for which the salt, ester or conjugate or its composition is administered when administered to the subject (including mammals, such as humans). The amount of a given composition described herein corresponding to such an amount may vary with various factors, such as a given agent, pharmaceutical formulation, route of administration, type of disease or disorder, identity of the subject (e.g., age, sex, weight) or the host being treated, etc.

[0025] As used herein, the term "treatment" refers to the administration of a compound or pharmaceutical composition for therapeutic purposes. "Treatment of a disorder" or for "therapeutic treatment" refers to the administration of treatment to a subject already suffering from a disease to improve the disease or one or more symptoms thereof, thereby improving the condition of the subject (e.g., by reducing one or more symptoms of inflammation). The compositions of the present disclosure may also be used as a primary preventive measure, i.e., to prevent a condition or reduce the risk of developing a condition. Prevention refers to preventive treatment of a subject who may not have fully developed a condition or disorder, but is susceptible to the condition or otherwise at risk of the condition. Therefore, in the claims and embodiments, the compositions and methods of the present disclosure may be used for therapeutic or preventive purposes.

[0026] As used herein, the terms "salt," "salt form," "conjugate," and "conjugate form" are interchangeable and are intended to include any compound formed when dephosphorylated cypridine is mixed with 1-hydroxy-2-naphthoic acid, 2-hydroxy-3-naphthoic acid, galactaric acid (mucic acid), glutamic acid, mandelic acid, naphthalene-2-sulfonic acid, camphorsulfonic acid, pantothenic acid, or isethionic acid. Without being bound by any particular theory, it is believed that the salts, esters, and conjugate forms of the compounds disclosed herein exhibit binding properties in addition to or in lieu of the ionic binding of conventional salts, which enhances their stability.

[0027] Other features and advantages of the present disclosure will be apparent from the following detailed description, examples, and claims.

[0028] Detailed Description

[0029] The present disclosure provides salt forms, esters, and conjugate forms of dephosphocyanidins useful in any therapy in which dephosphocyanidins can be used, pharmaceutical compositions comprising these salts, esters, and conjugates, and methods of producing the same.

[0030] The salts, esters and conjugates disclosed herein have the following general formula A:

[0031]

[0032] Wherein s / e / c in formula A represents salt, ester or conjugate.

[0033] In one non-limiting embodiment, the salt, ester or conjugate is prepared from a mixture of dephosphorylated cypridine and 1-hydroxy-2-naphthoic acid, 2-hydroxy-3-naphthoic acid, galactaric acid (mucic acid), glutamic acid, mandelic acid, naphthalene-2-sulfonic acid, camphorsulfonic acid, pantothenic acid or isethionic acid or an agent such as, but not limited to, chlorosulfonic acid, sulfur trioxide pyridine complex, sulfur trioxide N,N-dimethylformamide complex, sulfur trioxide triethylamine complex or sulfur trioxide trimethylamine complex.

[0034] A non-limiting example of a dephosphorylated cyperus salt, ester or conjugate of the present disclosure is dephosphorylated cyperus isethionate.

[0035] Another non-limiting example of a dephosphorylated cyprid salt, ester or conjugate of the present disclosure is dephosphorylated cyprid mucate.

[0036] In a non-limiting embodiment, the prepared salt, ester or conjugate is a zwitterion. A non-limiting example of a zwitterion of the present disclosure is dephosphorylated cyperidin-O-sulfate as depicted in Formula A1

[0037] .

[0038] Compared to dephosphocyanidin, the salts, esters and conjugates of the present disclosure exhibit improved stability, physical properties and / or handling characteristics. For example, dephosphocyanidin mucinate conjugates were demonstrated to be stable for at least 7 months.

[0039] In addition, studies in human subjects have shown that the salts and conjugates of the present disclosure exhibit strong sedative effects in patients without the hallucinogenic and / or anxiogenic effects commonly seen when dephospho-lamydopamine is administered at similar doses. Surprisingly, unlike the results reported for the administration of lamydopamine by Griffith et al. (Pharmaceutica Acta Helvetiae 2011 72(3) 175–184), 9 of 10 subjects administered dephospho-lamydopamine mucoacid conjugates at an equivalent effective dose of 2 mg dephospho-lamydopamine had no hallucinogenic effects, but rather strongly demonstrated an anxiolytic effect. These anxiolytic effects at such low doses without hallucinogenic effects were completely unexpected, especially in a patient population that typically undergoes blood draws within the same time period.

[0040] Also disclosed herein are pharmaceutical compositions comprising a dephosphorylated cyprid salt form, ester or conjugate form and a pharmaceutically acceptable excipient. The pharmaceutical compositions of the present disclosure exhibit improved stability, physical properties and / or handling characteristics compared to pharmaceutical compositions containing dephosphorylated cyprid. In addition, studies in patients have shown that the salts and conjugates of the present disclosure exhibit strong sedative effects in patients without the hallucinogenic and / or anxiogenic effects commonly seen when dephosphorylated cyprid is administered.

[0041] In general, the dosing regimens found to be useful for cypridine can also be used for the salts, esters and conjugates disclosed herein. In addition, microdosing, i.e., regular intake of dephosphorylated cypridine salts or conjugates disclosed herein at very low doses (ranging from 0.5-5 mg, depending on the condition to be treated), may also be an effective administration method with therapeutic potential.

[0042] Examples of pharmaceutically acceptable excipients include, but are not limited to, biocompatible carriers, adjuvants, additives, and diluents to obtain compositions that can be used as dosage forms. Examples of other excipients include colloidal silicon oxide, magnesium stearate, cellulose, sodium lauryl sulfate, and D&C Yellow #10.

[0043] The pharmaceutical compositions of the present disclosure may contain one or more solvents, diluents or other liquid carriers, dispersions or suspension aids, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders and lubricants suitable for the desired specific dosage form. Remington's Pharmaceutical Sciences, 18th edition, EW Martin (Mack Publishing Co., Easton, Pa., 1990) discloses various excipients and known preparation techniques for preparing pharmaceutical compositions. Unless any conventional excipient medium is incompatible with the compounds of the present invention, such as producing any undesirable biological effect or interacting with any other component of the pharmaceutical composition in other harmful ways, its use is considered to be within the scope of the present disclosure. Some examples of materials that can be used as pharmaceutically acceptable excipients include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; tragacanth powder; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; natural and synthetic phospholipids such as soybean and Egg yolk phospholipids, lecithin, hydrogenated soy lecithin, dimyristoyl lecithin, dipalmitoyl lecithin, distearoyl lecithin, dioleoyl lecithin, hydroxylated lecithin, lysophosphatidylcholine, cardiolipin, sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, distearoylphosphatidylethanolamine (DSPE) and its PEGylated esters (e.g., DSPE-PEG750 and DSPE-PEG2000), phosphatidic acid, phosphatidylglycerol, and phosphatidylserine; as well as hydroxypropyl-β-cyclodextrin and sulfonic acid-substituted cyclodextrins (e.g., CAPTISOL TM). Preferred commercial grade lecithins include those available under the trade names Phosal® or Phospholipon®, and include Phosal 53 MCT, Phosal 50 PG, Phosal 75 SA, Phospholipon 90H, Phospholipon 90G, and Phospholipon 90 NG; particularly preferred are soy phosphatidylcholine (SoyPC) and DSPE-PEG2000; buffers such as magnesium hydroxide and aluminum hydroxide may also be present in the composition at the discretion of the formulator; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; 5% dextrose solution and combinations of the above aqueous solutions; ethanol and phosphate buffered solutions, and other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as colorants, release agents, coating agents, sweeteners, flavoring agents and fragrances, preservatives, and antioxidants.

[0044] The pharmaceutical compositions of the present disclosure can be administered parenterally, orally, nasally, rectally, topically or buccally. The term "parenteral" as used herein refers to subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional or intracranial injection, and any suitable infusion technique.

[0045] When used in the methods and compositions of the present disclosure, the pharmaceutically acceptable dephosphorylated cyperine salt, ester or conjugate can be included in any suitable carrier material formulated for intravenous infusion in any suitable amount, and is generally present in an amount of 0.01-95% by weight based on the total weight of the composition. In a specific embodiment, the pharmaceutically acceptable dephosphorylated cyperine salt, ester or conjugate is present in an amount of 0.01-5% by weight based on the total weight of the composition. In some embodiments, an aqueous solution suitable for intravenous infusion comprising the pharmaceutically acceptable dephosphorylated cyperine salt, ester or conjugate can be formulated into a saline solution. The formulation of the infusion solution is well known to those skilled in the art of drug formulation. The formulation can be found in Remington: The Science and Practice of Pharmacy (23rd edition), AR Gennaro, ed., Lippincott Williams & Wilkins, 2000, and Encyclopedia of Pharmaceutical Technology, J. Swarbrick and JC Boylan, ed., 1988-1999, Marcel Dekker, New York. The composition for infusion can be provided in a unit dosage form (e.g., a single-dose ampoule) or a vial containing several doses of medicaments, to which a suitable preservative can be added. A solution of a pharmaceutically acceptable dephosphorylated cyperin salt, ester, or conjugate suitable for intravenous infusion can have a pH of about 3 to about 9. In addition, a solution of a pharmaceutically acceptable dephosphorylated cyperin salt, ester, or conjugate suitable for intravenous infusion can include a concentration of the pharmaceutically acceptable dephosphorylated cyperin salt, ester, or conjugate between about 0.1 mg / mL and about 50 mg / mL. In some embodiments, the aqueous solution has between about 1 mg / mL to about 15 mg / mL of any of the pharmaceutically acceptable salts, esters, or conjugates of dephosphocyanidin described herein. In specific embodiments, the aqueous solution has between about 0.1 mg / mL to about 1 mg / mL of any of the pharmaceutically acceptable salts, esters, or conjugates of dephosphocyanidin described herein.

[0046] Sterile injectable compositions may be solutions or suspensions in nontoxic parenterally acceptable diluents or solvents. Such solutions include, but are not limited to, 1,3-butanediol, mannitol, water, Ringer's solution, and isotonic sodium chloride solution. In addition, non-volatile oils are routinely used as solvents or suspending media (e.g., synthetic mono- or diglycerides). Fatty acids (e.g., but not limited to oleic acid and its glyceride derivatives) can be used to prepare injections, as can natural pharmaceutically acceptable oils (e.g., olive oil or castor oil) or polyoxyethylated versions thereof. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as, but not limited to, carboxymethylcellulose or similar dispersants. Other commonly used surfactants, such as, but not limited to, Tweens or Spans or other similar emulsifiers or bioavailability enhancers, which are commonly used to prepare pharmaceutically acceptable solid, liquid or other dosage forms, may also be used for formulation purposes.

[0047] Compositions for oral administration can be any oral acceptable dosage form, including capsules, tablets, emulsions and aqueous suspensions, dispersions and solutions. In the case of tablets, conventional excipients include, but are not limited to, lactose and corn starch. Lubricants are also usually added, such as, but not limited to, magnesium stearate. For oral administration in capsule form, useful diluents include, but are not limited to, lactose and dry corn starch. When aqueous suspensions or emulsions are administered orally, the active ingredient can be suspended or dissolved in an oil phase that is combined with an emulsifier or suspending agent. If desired, certain sweeteners, flavorings or coloring agents can be added.

[0048] Oral formulations such as capsules are preferably packaged in amber glass bottles secured with polypropylene caps with foam seal liners and maintained at 15-25°C in a sealed container in a dry location prior to administration.

[0049] In one non-limiting embodiment, the dephosphorylated cyperus salt, ester or conjugate is administered in the form of a nasal spray formulation.

[0050] In one non-limiting embodiment, the dephosphorylated cyprid salt, ester or conjugate is administered via a nasal spray sensor programmed time-release administration. A non-limiting device for such administration is described in PCT / US2021 / 028068, filed April 20, 2021, the teachings of which are incorporated herein by reference in their entirety.

[0051] In one non-limiting embodiment, the dephosphorylated cyperus salt, ester or conjugate is administered in the form of a nasal spray, wherein a therapeutically active amount of the dephosphorylated cyperus salt, ester or conjugate is dissolved or suspended in a solution or mixture of excipients (e.g., preservatives, viscosity modifiers, emulsifiers, buffers) in a non-pressurized dispenser that delivers a spray containing a metered dose of the active ingredient.

[0052] The dephosphorylated cyprid salts, esters and conjugates of the present disclosure and pharmaceutical compositions thereof can be used to treat or alleviate the symptoms of any disease or condition that can be treated with dephosphorylated cyprid or cyprid. Such methods include administering to a subject in need thereof a dephosphorylated cyprid salt, ester or conjugate of the present disclosure in an amount sufficient to treat or alleviate the symptoms of the disease or condition. Non-limiting examples of the disease or condition include nerve damage, neurodegenerative diseases, inflammatory conditions, chronic pain or psychological conditions. In certain embodiments, the disease or condition is an inflammatory condition (e.g., pulmonary inflammation, neuroinflammation, rheumatoid arthritis, atherosclerosis, psoriasis, type II diabetes, inflammatory bowel disease, Crohn's disease, multiple sclerosis and / or sepsis). In specific embodiments, the inflammatory condition is chronic obstructive pulmonary disease (COPD) or Alzheimer's disease. In certain embodiments, the disease or condition is nerve damage (e.g., stroke, traumatic brain injury or spinal cord injury). In some embodiments, the disease or condition is chronic pain (e.g., pain caused by postoperative pain, tension headaches, chronic low back pain, fibromyalgia, kidney disease, multiple sclerosis, herpes zoster, complex regional pain syndrome, headaches, or sciatica). In specific embodiments, the chronic pain condition is caused by trigeminal autonomic headaches (e.g., episodic and chronic cluster headaches (CH), episodic and chronic paroxysmal hemicrania (PH), and short-term unilateral neuralgiform headache attacks with conjunctival injection and tearing (SUNCT)). In some embodiments, the trigeminal autonomic headache is episodic or chronic CH. In certain embodiments, the condition is a psychological condition (e.g., depression, anxiety, addiction, post-traumatic stress disorder, eating disorders, selective mutism, or obsessive-compulsive behavior). In specific embodiments, the psychological condition is depression or anxiety.

[0053] The following examples more fully demonstrate the features and advantages of the dephosphorylated cyperus salts, esters and conjugates of the present disclosure and compositions prepared therefrom, which are provided for illustrative purposes and should not be construed as limiting the present disclosure in any manner.

[0054] Example

[0055] Preparation of dephosphorylated cyperus salts, esters and conjugates

[0056] Material:

[0057] The following acids were used in the preparation of novel dephosphorylated cyprid salts and conjugates:

[0058] 1-Hydroxy-2-naphthoic acid

[0059] 3-Hydroxy-2-naphthoic acid

[0060] Galactaric acid (mucic acid)

[0061] L-Glutamic Acid

[0062] Mandelic acid (R-2-hydroxy-2-phenylacetic acid)

[0063] Pantothenic acid

[0064] Isethionic acid [2-Hydroxyethylsulfonic acid]

[0065] Naphthalene-2-sulfonic acid

[0066] (1S)-(+)-10-Camphorsulfonic acid

[0067] In addition, a dephosphorylated cypridine ester, dephosphorylated cypridine-O-sulfate, was prepared, which, like cypridine, is a zwitterion. Dephosphorylated cypridine-O-sulfate can be prepared by reacting dephosphorylated cypridine with a variety of reagents, such as, but not limited to, chlorosulfonic acid, sulfur trioxide pyridine complex, sulfur trioxide N,N-dimethylformamide complex, sulfur trioxide triethylamine complex, or sulfur trioxide trimethylamine complex.

[0068] General procedure:

[0069] To prepare salts, esters and conjugates, equimolar or greater amounts of dephosphorylated cyprid and corresponding acids or other reagents are used. Various solvents are used to prepare dephosphorylated cyprid salts, esters and conjugates, and then the salt, ester or conjugate product is recrystallized; typical solvents used are, for example, methanol, ethanol, tetrahydrofuran, acetone, ethyl acetate and ether.

[0070] Embodiment 1:

[0071] 204.2 mg (1.0 mmol) of dephosphocyanidin and the corresponding acid (1.0 mmol) selected from 1-hydroxy-2-naphthoic acid, 2-hydroxy-3-naphthoic acid, galactaric acid (mucoic acid), glutamic acid, mandelic acid, naphthalene-2-sulfonic acid, camphorsulfonic acid were weighed into an 8 ml vial and then 2.0 ml of methanol was added while stirring with a magnetic bar. Precipitates formed immediately with mucoic acid and 1-hydroxy-2-naphthoic acid. All other salts or conjugates formed clear solutions except for glutamic acid which formed a sticky substance.

[0072] After dilution with about 4 mL of ethyl acetate, the precipitate was collected on a sintered glass frit. The collected product was washed sequentially with ethyl acetate and diethyl ether, and then dried under high vacuum.

[0073] In reactions where no precipitate formed, the clear solution was transferred to a small round bottom flask. The solvent was removed using a rotary evaporator; a foam formed when a high vacuum was applied. The foam was then treated with a combination of solvents including, but not limited to, ethyl acetate, acetone, tetrahydrofuran, and diethyl ether to precipitate the product.

[0074] Embodiment 2:

[0075] The pantothenate salt of dephosphocyanidin was prepared as follows. 476.6 mg (1.0 mmol) of calcium pantothenate was dissolved in 2.0 ml of deionized water and treated with 2.0 ml of 1N HCl; a clear solution was obtained. This solution was added to a stirred solution of 408.4 mg (2.0 mmol) of dephosphocyanidin in 2.0 ml of methanol. After stirring at room temperature for 2 hours, the solvent was removed and dried under high vacuum to obtain a white solid containing the pantothenate salt of dephosphocyanidin and 1 mmol of calcium chloride.

[0076] Embodiment 3:

[0077] The isethionate salt of dephosphocyanidin was prepared as described here. 145.4 mg (1.0 mmol) of sodium isethionate was dissolved in 2.0 mL of deionized water and treated with 1.0 mL of 1N HCl; a clear solution was obtained. This solution was added to a stirred solution of 204.2 mg (1.0 mmol) of dephosphocyanidin in 3.0 mL of methanol. After stirring at room temperature for 2 hours, the solvent was removed and dried under high vacuum to obtain a white solid containing the isethionate salt of dephosphocyanidin and 1 mmol of sodium chloride.

[0078] Embodiment 4:

[0079] Dephosphocyanidin-O-sulfate is prepared, for example, as follows: a solution of 204.0 mg (1.0 mmol) of dephosphocyanidin in 5.0 mL of pyridine is treated with 318.3 mg (2.0 mmol) of sulfur trioxide pyridine complex. The reaction is continued overnight, and then the solvent is removed under high vacuum. Cold water is added to the residue, and the insoluble product is collected on a sintered glass frit. The product is washed with cold water and methanol and dried under high vacuum. Recrystallization from methanol gives the pure product.

[0080] Example 5: Purity Assessment

[0081] The purity of the dephosphocyanin salts, esters and conjugates described below was assessed by high pressure liquid chromatography (HPLC).

[0082] Salt / ester / conjugate Molecular weight Dephosphocyanidin mucate 414.30 Dephosphorylation of chlorophyll OH naphthoate 392.30 Dephosphorylation of cyperus mandelate 356.30 <![CDATA[Dephosphorylated psilocin pantothenate 1 > 534.38 Dephosphocyanidin O-sulfate 284.2 Dephosphocyanine 204.27

[0083] 1 Contains approximately 1 mol • CaCl2

[0084] Reverse phase HPLC was used to determine the chromatographic purity, content determination and retention time identification of the synthesized dephosphorylated cyprid salts, esters and conjugates. Chromatographic separation was performed on a 4.6 x 250 mm, 5 µm Zorbax SB-Phenyl column using gradient elution and UV detection at 220 nm. Diode array detection was also used to identify and analyze the dephosphorylated cyprid salts, esters and conjugates in the range of 200 nm to 400 nm. The chromatographic conditions used are shown in Table 1. A storage solution of the sample was prepared in methanol at a concentration of about 0.5 mg / mL dephosphorylated cyprid. The sample was diluted five times in dilute acetic acid (0.2% v / v) to make the concentration of dephosphorylated cyprid in 80:20 dilute acetic acid: methanol to be about 0.1 mg / mL. Purity values ​​were determined using peak area percentage. Dephosphorylated cyprid base was used as an external reference standard to calculate the content determination values ​​of the dephosphorylated cyprid salts, esters and conjugates. The assay values ​​are on an "as is" basis, without regard to moisture, residual solvents or inorganic impurities. The retention time and UV spectrum of dephosphocyanidin O-sulfate did not yield a positive identification of dephosphocyanidin. The results are shown in Table 2.

[0085] Table 1. HPLC chromatographic conditions

[0086]

[0087] Table 2. Results of assay, chromatographic purity and identification

[0088]

[0089] 1 Contains approximately 1 mol • CaCl2, with a corrected assay value of 103.8%

[0090] 2 O-sulfate is an ester on the phenolic hydroxyl group

[0091] 3 Does not match the positive identification of dephosphocyanin (UV spectrum and retention time)

[0092] Example 6: Stability study of dephosphorylated cyperus mucate

[0093] Additional stability studies were performed on dephosphocyanidin mucate (also known as dephosphocyanidin mucate or conjugate or dephosphocyanidin galactarate or conjugate) capsules.

[0094] Appearance was assessed by viewing the sample on a watch glass against a black and white background.

[0095] The assay conditions for the test / method TM-022-0392 used to evaluate stability and impurities are as follows:

[0096] Mobile phase: MPA: 945 mL water, 5 mL H3PO4, pH adjusted to 5.7 with NaOH; MPB: acetonitrile

[0097] Diluent: 80:20 water:MeOH

[0098] Standard: 0.4 mg / mL dephosphocyanidin mucate RS in diluent

[0099] Sample: 0.4 mg / mL dephosphocyanidin mucate API in diluent

[0100] The HPLC conditions are depicted in Table 3.

[0101] Table 3

[0102]

[0103] Additional balancing may be required

[0104] System Kit: RSD NMT 2.0% (5 injections), Tailing NMT 2.0, Plate Count NLT 10000

[0105] Calculations were performed for the external standards in brackets.

[0106] The identity of dephosphocyanidin mucate was confirmed by ultraviolet (UV) evaluation, where the UV spectrum of the major peak in the standard matched that of the major peak in the sample, and by retention time, where the retention time ratio was 0.98–1.02.

[0107] The results are depicted in Table 4 below.

[0108] Table 4

[0109]

[0110] Example 7: A Phase I, Open-label, Single-treatment, Single-dose, Single-cycle, Pharmacokinetic Study of Hard Gelatin Capsules Containing Dephosphocyanidin as a Mucic Acid Conjugate

[0111] For this study, a tryptamine derivative, dephosphocyanidin mucate, was administered as a white to light brown solid crystalline powder. The molecular weight of this compound is 414.41 g / mol. In terms of equivalents, 4.05 mg of dephosphocyanidin mucate is equivalent to 2 mg of dephosphocyanidin. In humans, the mucate is enzymatically cleaved during metabolism in vivo to produce dephosphocyanidin, which is an agonist of various serotonin receptors including the 5-HT2A receptor, which is the basis for the hallucinogenic and therapeutic effects of dephosphocyanidin (Cao et al., Science (New York, NY) 2022 375(6579):403–411; Lowe et al., Molecules 2021 26(10), 2948). For this clinical study, dephosphocyanidin was provided as white / opaque hard gelatin capsules containing 4.05 mg of dephosphocyanidin mucoconjugate, equivalent to 2 mg of dephosphocyanidin. 2 One capsule was administered orally with water to healthy subjects (males at least 50 kg and females at least 45 kg), non-smokers or those who had stopped smoking 24 hours prior to dosing. The objectives of the study were to evaluate the bioavailability and pharmacokinetic parameters of the mucoid acid conjugate by measuring plasma concentrations of dephosphocyanidin and performing calculations based on these measurements.

[0112] Blood (8 mL) was collected in K3 EDTA blood collection tubes under a sodium lamp before dosing (-1.00) and at 0.25, 0.50, 0.75, 1.00, 1.50, 2.00, 3.00, 5.00, 8.00, 12.00, 16.00, and 24.00 h after dosing.

[0113] A total of 13 blood samples were drawn during the study period.

[0114] In addition, a psychiatrist independent of the study's principal investigator assessed each subject's "mood" and gave each subject a Mini-Mental State Examination (MMSE) test to determine if the subject's score decreased. The MMSE test is an 11-question measure that tests five areas of cognitive function: orientation, registration, attention and calculation, recall, and language. When hallucinogenic substances are administered, MMSE scores are expected to drop below 25. The results are depicted in Table 5 below.

[0115] Table 5

[0116]

[0117] Surprisingly, and in contrast to the results reported for example by Griffith et al. (Pharmaceutica Acta Helvetiae 2011 72(3) 175–184) with cyperidin administration, 9 of 10 subjects administered dephospho-cyperidin mucoacid conjugates had no hallucinogenic effects but strongly suggested anxiolytic effects. These anxiolytic effects at such low doses without hallucinogenic effects were completely unexpected, particularly in a patient population that typically undergoes blood draws within the same time frame.

Claims

1. A dephosphorylated cyperus salt, ester or conjugate of formula A, ， Where s / e / c represents salt, ester or conjugate, It is prepared from a mixture of dephosphorylated cypridine and 1-hydroxy-2-naphthoic acid, 2-hydroxy-3-naphthoic acid, galactaric acid (mucinic acid), glutamic acid, mandelic acid, naphthalene-2-sulfonic acid, camphorsulfonic acid, pantothenic acid, isethionic acid, chlorosulfonic acid, sulfur trioxide pyridine complex, sulfur trioxide N,N-dimethylformamide complex, sulfur trioxide triethylamine complex or sulfur trioxide trimethylamine complex.

2. The dephosphorylated cypridine conjugate according to claim 1, which is prepared from a mixture of dephosphorylated cypridine and galactaric acid (mucic acid).

3. The dephosphorylated cyperus ester according to claim 1, which is depicted in formula A1, It is prepared from a mixture of dephosphorylated cyperidin and sulfur trioxide pyridine complex.

4. A pharmaceutical composition comprising the dephosphorylated cyperus salt, ester or conjugate according to any one of claims 1, 2 or 3 and one or more pharmaceutically acceptable excipients.

5. A method for producing a dephosphorylated cypridine salt, ester or conjugate according to claim 1, the method comprising mixing dephosphorylated cypridine with 1-hydroxy-2-naphthoic acid, 2-hydroxy-3-naphthoic acid, galactaric acid (mucic acid), glutamic acid, mandelic acid, naphthalene-2-sulfonic acid, camphorsulfonic acid, pantothenic acid, isethionic acid, chlorosulfonic acid, sulfur trioxide pyridine complex, sulfur trioxide N,N-dimethylformamide complex, sulfur trioxide triethylamine complex or sulfur trioxide trimethylamine complex.

6. A method for producing the dephosphorylated cypridine conjugate according to claim 2, comprising mixing dephosphorylated cypridine with galactaric acid (mucic acid).

7. A method for producing the dephosphorylated cypridine ester according to claim 3, comprising mixing dephosphorylated cypridine with a sulfur trioxide pyridine complex.

8. A method for treating a disease or condition treatable with dephosphocyanidin or cyanidin in a subject, the method comprising administering to the subject a pharmaceutical composition according to claim 4.

9. The method of claim 8, wherein the disease or disorder is nerve damage, a neurodegenerative disease, an inflammatory disorder, chronic pain, or a psychological disorder.

10. The method of claim 8, wherein the disease or condition is chronic cluster headache.

11. A method of reducing adverse effects of administering dephosphocyanin in a subject, the method comprising administering the pharmaceutical composition according to claim 4 to the subject.

12. A method of using dephosphocyanin to reduce anxiety in a subject without hallucinogenic effects, the method comprising administering to the subject a compound according to claim 2.

13. Use of the pharmaceutical composition according to claim 4, for treating a disease or condition treatable with dephosphocyanidin or cyanidin in a subject.

14. The use according to claim 13, wherein the disease or disorder is nerve damage, a neurodegenerative disease, an inflammatory disorder, chronic pain or a psychological disorder.

15. The use according to claim 13, wherein the disease or disorder is chronic cluster headache.

16. Use of the pharmaceutical composition according to claim 4, for alleviating adverse reactions of dephosphocyanin treatment in a subject.

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