Aspirin simulant antioxidant for treating amyotrophic lateral sclerosis

Abstract

The invention discloses an aspirin simulant antioxidant for treating amyotrophic lateral sclerosis, which is an active pharmaceutical ingredient selected from DPL-001, DPL-0012, DPL-003 and DPL-004, or a pharmaceutically acceptable salt of the active pharmaceutical ingredient. According to the invention, ALS is treated by enteric capsules and / or intravenous drip with a novel aspirin mimetic antioxidant. When the capsule is used by six patients with ALS in the same situation for six months, the progress of the disease can be obviously slowed down by orally taking one capsule every day, and even the ALS can be slightly reversed under some conditions. Compared with edaravone, the curative effect of a new drug (DPL-002) is remarkable, and the curative effect is 67%.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, in particular to an aspirin mimetic antioxidant for treating amyotrophic lateral sclerosis. Background Art

[0002] There is a finely regulated redox system in living organisms. In a healthy human body, any harmful oxidants from endogenous and exogenous sources can be neutralized promptly and effectively by reductases to protect important biomolecules such as proteins, DNA, and lipids from oxidation. However, in an unbalanced situation, there are more harmful oxidants than the processing capacity of reductases, which is called a state of oxidative stress. Excessive oxidants can oxidize important biomolecules such as proteins, DNA, and lipids, causing dysfunction or malfunction of these biomolecules, gradually leading to disease and even death. These harmful oxidants are basically reactive oxygen species (ROS) and reactive nitrogen species (RNS). Among them, most are free radicals. Free radicals tend to acquire an electron or a hydrogen radical to become a stable substance, and this process is called free radical oxidation.

[0003] There are three sources of free radicals in the human body. Internal sources include mitochondria, inflammation, exercise, xanthine oxidase, peroxisomes, phagocytes, etc. Free radicals from internal sources are generated from normal metabolic cycles. Some of them are used by the immune system to fight against bacterial or viral invasion. Their functions also include redox signaling, clearing dead cells, activating and regulating some important life processes, etc. In general, free radicals from internal sources are usually controlled by the redox system. External sources include cigarette smoke, alcoholism, toxins, certain drugs, ozone, ultraviolet rays, radiation, pesticides, herbicides, environmental pollutants, etc. The third source is related to physiological factors such as stress, emotions, and disease conditions. Free radicals from the latter two sources are usually an additional burden on the redox system, and they are the root cause of oxidative stress that causes health problems.

[0004] Although the pathogenesis and pathology of many major diseases remain elusive, free radicals are closely related to the etiology and pathology of many major diseases, including Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), atherosclerosis, cardiovascular disease, kidney disease, nephropathy, hypertension, hyperlipidemia, rheumatoid arthritis, lupus, multiple sclerosis (MS), gout, inflammation, pain, aging, stroke, depression, acne, cataracts, glaucoma, age-related macular degeneration, cancer and HIV. Antioxidants that can fight free radicals have attracted widespread attention in the past few decades. The theory of free radicals and antioxidants related to human health has been widely accepted worldwide. This theory was first proposed by Dr. Denham Harman in the 1950s. A large number of research papers in this field have been published in a wide range of scientific journals, and this trend continues. However, there are still very few antioxidant drugs approved for the treatment of certain major diseases.

[0005] Antioxidants are compounds that can inactivate active oxidants by being oxidized themselves. Organic antioxidants are generally considered reducing agents due to their ease of oxidation and are divided into three types: aniline-like compounds including indoles, phenols and polyphenolic compounds, thiol-containing compounds such as cysteine ​​and lipoic acid, and selenol-containing compounds.

[0006] Natural sources of antioxidants include fruits, vegetables, tea (EGCG), red wine (tannins), and other dietary habits. Plant polyphenols, flavonoids, beta-carotene, vitamin A, vitamin C, vitamin E, salicylic acid, hemp, curcumin, lipoic acid, dithiocarboxylic acid, omega-3, oleocanthal, glutathione, and melatonin are examples of natural antioxidants.

[0007] So far, synthetic organic antioxidants have been much less explored than natural antioxidants. BHT (2,6-di-tert-butyl-4-methylphenol) is an example of a synthetic organic antioxidant, which is widely used to store some organic solvents, including THF and diethyl ether, to protect them from oxidation by air. Acetaminophen (Tylenol), oltipraz, DTT (dithiothreitol), succinylbutrol, and probucol are other examples of synthetic antioxidants. There are some obvious advantages of synthetic organic antioxidants, their molecular structure can be adjusted to improve bioavailability, reduce toxicity, enhance free radical scavenging ability, etc. Synthetic antioxidants will certainly play an important role in the intervention of major diseases related to oxidative stress.

[0008] Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease. The neurological disorder affects the motor neurons of the brain and spinal cord. Motor neurons are nerve cells that control voluntary muscle movement and breathing. In ALS, motor neurons gradually lose function and die. The patient inevitably becomes paralyzed and eventually dies from respiratory failure. Most patients die within 3-5 years of the first onset of symptoms. However, 10% survive for a decade or more. ALS is a rare disease, with an incidence of about two people diagnosed with ALS per 100,000 people each year. The cause of ALS is unknown, although 2-5% of ALS cases are inherited from the family.

[0009] Although the FDA has approved some drugs / therapies to treat ALS, there is currently no effective cure for ALS. Approved drugs / therapies may slow the progression of the disease, prolong survival, or improve quality of life. But they cannot reverse the progression of the disease or stop it from progressing, nor can they repair damage to motor neurons. Researchers continue to work on developing new drugs to better intervene in ALS.

[0010] Therefore, the new drug of the present invention can provide a new option for patients with amyotrophic lateral sclerosis (ALS), thereby better meeting unmet medical needs. Summary of the invention

[0011] The purpose of the present invention is to provide an aspirin mimetic antioxidant for treating amyotrophic lateral sclerosis to solve the problems in the prior art.

[0012] To achieve the above object, the present invention provides the following technical solutions: an aspirin mimetic antioxidant for treating amyotrophic lateral sclerosis, an active pharmaceutical ingredient selected from DPL-001, DPL-0012, DPL-003, DPL-004, or a pharmaceutically acceptable salt thereof;

[0013] The molecular structure of DPL-001 is as follows:

[0014]

[0015] The molecular structure of DPL-002 is as follows:

[0016]

[0017] The molecular structure of DPL-003 is as follows:

[0018]

[0019] The molecular structure of DPL-004 is as follows:

[0020]

[0021] An intravenous drip solution, characterized in that it comprises the compound, a pharmaceutically acceptable carrier and optionally one or more pharmaceutical excipients.

[0022] Preferably, the pharmaceutically acceptable carrier is water, and the infusion solution further comprises a pharmaceutical excipient selected from sodium chloride, potassium chloride, sodium bisulfite and a combination thereof.

[0023] Preferably, the dosage range of the active pharmaceutical ingredient is 5 mg to 1000 mg per day, or from 0.1 mg / kg to 20 mg / kg body weight, or from 0.47 μmol / kg to 94 μmol / kg body weight.

[0024] Preferably, the dosage is administered to the subject at a frequency of once to five times per day.

[0025] A capsule comprises the compound, a pharmaceutically acceptable carrier and optionally one or more pharmaceutical excipients.

[0026] Preferably, the pharmaceutically acceptable carrier is an enteric-coated capsule, wherein the enteric-coated capsule further comprises a pharmaceutical excipient selected from starch powder, purified water, sodium chloride, potassium chloride, dicalcium phosphate, magnesium stearate, sodium carboxymethylcellulose and a combination thereof.

[0027] Preferably, the dosage of the active pharmaceutical ingredient is 10 mg to 2800 mg per day, or 0.2 mg / kg to 56 mg / kg body weight, or 0.94 μmol / kg to 263.2 μmol / kg body weight.

[0028] Preferably, the frequency of administration to the subject is from once a day to five times a day.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] 1. Treat ALS with a new aspirin-mimetic antioxidant via enteric-coated capsules and / or intravenous infusion. During the 6-month compassionate use of 6 ALS patients, taking one capsule orally per day significantly slowed the progression of the disease and even slightly reversed ALS in some cases. Compared with edaravone, the new drug (DPL-002) has a significant efficacy of 67%. Given that there is currently no effective treatment for ALS, the current medical needs of this incurable motor neuron disease can be met by the new drug (DPL-002). BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0032] Figure 1 It is an existing aniline antioxidant.

[0033] Figure 2 It is an antioxidant of some phenolic types.

[0034] Figure 3 It is an example of a synthetic phenolic antioxidant.

[0035] Figure 4 It is leonurine, catechins and cannabinoids that act as phenolic antioxidants.

[0036] Figure 5 They are aspirin, salicylic acid and their derivatives.

[0037] Figure 6 It is a kind of antioxidant containing thiol.

[0038] Figure 7 It is a dithiol-containing antioxidant.

[0039] Figure 8 It is a dithiol-containing antioxidant.

[0040] Fig. 9 They are thioaspirin, thimerosal and thiosalicylic acid.

[0041] Fig.10 It is a new drug that selectively forms stable chelates with heavy metals.

[0042] Fig.11 The invention discloses some novel aspirin mimetics and intermediates thereof. DETAILED DESCRIPTION

[0043] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. Therefore, the following detailed description of the embodiments of the present invention provided in the drawings is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention.

[0044] In an embodiment of the present invention, an aspirin mimetic antioxidant for treating amyotrophic lateral sclerosis, an active pharmaceutical ingredient selected from DPL-001, DPL-0012, DPL-003, DPL-004, or a pharmaceutically acceptable salt thereof;

[0045] The molecular structure of DPL-001 is as follows:

[0046]

[0047] The molecular structure of DPL-002 is as follows:

[0048]

[0049] The molecular structure of DPL-003 is as follows:

[0050]

[0051] The molecular structure of DPL-004 is as follows:

[0052]

[0053] An intravenous drip solution, characterized in that it comprises the compound, a pharmaceutically acceptable carrier and optionally one or more pharmaceutical excipients.

[0054] Preferably, the pharmaceutically acceptable carrier is water, and the infusion solution further comprises a pharmaceutical excipient selected from sodium chloride, potassium chloride, sodium bisulfite and a combination thereof.

[0055] Preferably, the dosage range of the active pharmaceutical ingredient is 5 mg to 1000 mg per day, or from 0.1 mg / kg to 20 mg / kg body weight, or from 0.47 μmol / kg to 94 μmol / kg body weight.

[0056] Preferably, the dosage is administered to the subject at a frequency of 1 to 5 times per day.

[0057] A capsule comprises the compound, a pharmaceutically acceptable carrier and optionally one or more pharmaceutical excipients.

[0058] Preferably, the pharmaceutically acceptable carrier is an enteric-coated capsule, wherein the enteric-coated capsule further comprises a pharmaceutical excipient selected from starch powder, purified water, sodium chloride, potassium chloride, dicalcium phosphate, magnesium stearate, sodium carboxymethylcellulose and a combination thereof.

[0059] Preferably, the dosage of the active pharmaceutical ingredient is 10 mg to 2800 mg per day, or 0.2 mg / kg to 56 mg / kg body weight, or 0.94 μmol / kg to 263.2 μmol / kg body weight.

[0060] Preferably, the frequency of administration to the subject is from once a day to five times a day.

[0061] ALS is treated with a novel aspirin-mimetic antioxidant via enteric-coated capsules and / or intravenous infusion. During 6 months of compassionate use in 6 ALS patients, one capsule taken orally daily significantly slowed disease progression and even slightly reversed ALS in some cases. The new drug (DPL-002) had a significant 67% efficacy compared to edaravone. Given that there are currently no effective treatments for ALS, the medical needs of this currently incurable motor neuron disease could be met by the new drug (DPL-002).

[0062] 1. Known natural antioxidants and synthetic antioxidants

[0063] Organic antioxidants are usually a nitrogen-, oxygen-, sulfur-, or selenium-containing compound. Nitrogen-containing molecules are usually aniline-like (including aniline-like) because anilines are more easily oxidized than amines. Edaravone, pyrenoxine, falconol, melatonin, and bendazac are examples of aniline antioxidant compounds. Figure 1 The other four are chemically synthesized. Melatonin is found in the human body and is secreted by the pineal gland. Melatonin is also known for its hormonal function. Pirenoxine is also a phenolic antioxidant because it contains phenol. Edaravone has been approved for marketing in Japan, China and the United States.

[0064] Oxygenated antioxidants are usually phenols, because phenols are more easily oxidized than alkyl alcohols. Acetaminophen (Tylenol), resveratrol, vitamin C, luteolin (a type of flavonoid), and vitamin E are examples of phenolic antioxidants ( Figure 2 ). All of these, except acetaminophen, are natural antioxidants. In the body, acetaminophen produces 4-aminophenol, which is a phenolic and aniline antioxidant.

[0065] An interesting example of a synthetic phenolic antioxidant is succinylbutyrol (AGI-1067) from AtheroGenics, which has a similar structure to BHT, such as Figure 3 As shown. Both are phenolic antioxidants. The steric hindrance caused by these tert-butyl groups may have a subtle effect on pharmacodynamics. In fact, steric hindrance was taken into consideration when designing some of the new aspirin mimetics in the present invention.

[0066] Leonurine ( Figure 4 ) was originally isolated from herbal extracts and is now chemically synthesized and can be considered a phenolic antioxidant compound. Leonurine has been reported to reduce arterial plaque and fibrosis in rabbits in preclinical tests. However, there are no reports on whether clinical trials have been conducted. Among the many naturally occurring antioxidants, catechins, cannabinoids ( Figure 4 ), curcumin, flavonoids, and tannins are good examples of phenolic antioxidants.

[0067] Aspirin has been used for indications for more than 120 years and is still commonly used around the world. In the body, aspirin is converted into salicylic acid, a phenolic antioxidant. Therefore, aspirin can be considered a prodrug of phenolic antioxidants. NOSH-aspirin, known as super aspirin, is a derivative of salicylic acid. It contains both a nitric oxide-releasing group and a hydrogen sulfide-releasing group ( Figure 5 ). Preclinical studies have reported that NOSH-aspirin is used in preclinical studies to prevent cancer, prevent inflammation, and prevent heart attacks. Similarly, Figure 5 NCX-4016 (nitroaspirin) in the drug is a derivative of aspirin that has also been reported to prevent heart attacks in preclinical studies.

[0068] Cysteine ​​is a sulfur-containing, naturally occurring amino acid known for its function in biological redox systems. Its oxidized form, cystine, plays an important role in the higher-order structure / architecture of proteins (enzymes). Figure 6 Glutathione, egg mercaptan and NAC (N-acetylcysteine) in glutathione are monothiol antioxidants. Glutathione participates in the enzyme-catalyzed cycle of the redox system in all tissues and organs. In healthy cells and tissues, the ratio of the disulfide bond type (GSSG) to the reduced type (GSH) is less than 1:9. An increase in the ratio of GSSG to GSH is an indicator of oxidative stress.

[0069] Figure 7 Alpha lipoic acid (ALA), DTT (dithiothreitol), dithiothiolane, dithiomethoxazole and oltipraz in are dithiol antioxidants. When the dithiol compound forms a five-membered ring through a disulfide bond, it is a stronger free radical scavenger than the corresponding monothiol compound. Alpha lipoic acid usually exists in the disulfide form (oxidized form) because the reduced form is easily oxidized by air.

[0070] Figure 8 In the chemistry of cyclization, DTT (dithiothreitol) is often used as a reagent to break the disulfide bonds of the substrate and form a new six-membered disulfide bond by itself (WW Cleland, Biochemistry, v.3, 480, 1964). Obviously, the driving force is the cyclization reaction itself, through the formation of a new disulfide bond. However, due to toxicity issues, neither DTT nor DTE (dithioerythritol) have been used as drugs. Some other interesting dithiol-containing antioxidants. The sulfur atom is in the allylic position, and the sulfur atom in the allylic position is activated, which helps to form disulfide bonds.

[0071] 2. Sulfo-aspirin and derivatives

[0072] Aspirin is used for many indications. Its molecular structure is considered to be a key scaffold for the use of new drugs. Some aspirin analogs have been reported, such as thioaspirin and thimerosal ( Fig. 9 Thimerosal is toxic due to its ethylmercury moiety and is approved only for topical antibacterial / antifungal agents such as antiseptics, nasal sprays, eye drops, etc. No preclinical studies have been reported on thioaspirin. Both thioaspirin and thiomerosal can produce thiosalicylic acid ( Fig. 9 ). Sulfosalicylic acid is a thiophenol antioxidant.

[0073] 3. Clinical trial results of several antioxidant drug candidates

[0074] Many clinical trials have investigated antioxidants, most of which are natural antioxidants, such as resveratrol, vitamin A, vitamin C, vitamin E, melatonin, beta-carotene, flavonoids, curcumin, etc., in the intervention of various diseases, such as neurodegenerative diseases, cancer, diabetes, inflammation, atherosclerosis, aging, etc. No definite benefits, marginal benefits, or controversial results have been obtained so far. In some randomized controlled clinical trials, vitamin A, vitamin E, or beta-carotene caused higher side effects, such as higher mortality or higher risk of certain cancers. AGI-1067( Figure 3 ) is an investigational drug for type II diabetes, inflammation, and cardiovascular disease indications. However, it failed to show efficacy in a Phase III clinical trial in 2008. Figure 1 ) has been approved in Japan, China, and the United States for acute ischemic stroke (AIS) and / or ALS. However, edaravone is a compound discovered in the 1950s with weak antioxidant capacity.

[0075] 4. Four FDA-approved drugs for the treatment of ALS

[0076] Although there is no known cure, the FDA has approved four drugs for the treatment of ALS. Riluzole was approved by the FDA in 1995 and can extend the life of patients to 2-3 months. Edaravone was approved by the FDA in 2017. In a 6-month clinical trial, it can slow the progression of early ALS by 33.3% compared with placebo. Notably, Edaravone (Radicava) slows the progression of ALS at all stages. Relyvrio (sodium phenylbutyrate / taurine diol) was approved by the FDA in 2022 and can extend the life of patients in the early stages of ALS by 5.5 months.

[0077] Tofersen, an mRNA drug (MW 7128 Daltons) approved by the FDA in 2023 for the treatment of familial hereditary ALS (genetic) by intrathecal injection. SOD1-ALS patients account for 2% of the total number of ALS patients. Tofersen reduces NfL but does not slow the progression of ALS.

[0078] The shortcomings of currently known antioxidants and their solutions

[0079] All antioxidants used in preclinical and clinical studies so far are not good terminators of free radicals. When these antioxidants extinguish primary free radicals, they themselves become new free radicals, called secondary free radicals. Secondary free radicals are less reactive compared to primary free radicals (Scheme 1). Compared to primary free radicals, secondary free radicals are less reactive and have a longer lifetime, thus traveling a longer distance within the cell / biological system. Based on the molecular structure of the antioxidants, antioxidant-derived secondary free radicals can reach some critical parts and damage DNA, lipids, and proteins through free radical oxidation. Given their lower reactivity, why do secondary free radicals still cause damage? There are two explanations. First, despite their lower reactivity compared to primary free radicals, they are still strong enough to directly oxidize some fragile DNA, lipids, and proteins, etc. Second, secondary free radicals may be too weak to directly oxidize biomolecules such as DNA, lipids, and proteins, but they can easily oxidize DNA, lipids, and proteins, etc. when some metal cations (Fe, Cu, etc) are present in the body as catalysts for electron transfer.

[0080]

[0081] Scheme 1. Free radical propagation

[0082] For example, vitamin E is a well-known strong antioxidant. However, when it quenches a free radical, it itself becomes a free radical (Scheme 2), which is called a secondary free radical in the process of propagation. The free radical form of vitamin E is a pro-oxidant because it can still oxidize some other molecules by acquiring electrons or hydrogen radicals. Therefore, the free radical form of vitamin E can still be harmful, especially in the presence of some metal cations (Fe, Cu, etc.) as electron transfer catalysts. Therefore, vitamin E is not a very good free radical scavenger in this regard. The same conclusion can be drawn if the situation of other antioxidants is considered.

[0083] This can well explain the results of some randomized controlled clinical trials on antioxidants. In the 2012 ADCS randomized controlled trial, Douglas R. Galasko et al. found that Alzheimer's patients treated with a combination of antioxidants (vitamin E / C / ALA) showed a decrease in cerebrospinal fluid biomarkers of oxidative stress, but accelerated cognitive decline, and no changes in cerebrospinal fluid pathological markers. Oxidative stress is reduced because antioxidants extinguish more active primary free radicals, which themselves become less active secondary free radicals. Since the activity of free radicals in the body is reduced, the measured oxidative stress will definitely decrease. Since secondary free radicals can reach certain key sites over a longer distance, they can cause damage directly through non-catalytic oxidation, or through catalytic oxidation of certain metal cations (Fe, Cu, etc.) as catalysts for electron transfer, which leads to accelerated cognitive decline. The results of this randomized controlled trial disappointed the researchers, but on the other hand, it also clearly proved that free radicals are one of the causes of AD, if not the only cause.

[0084]

[0085] Scheme 2. Tocopherol (α-vitamin E) quenches free radicals

[0086] Some other negative results have also been observed in randomized controlled clinical trials for cancer prevention (Table 1), such as the ATBC randomized controlled trial by Olli P. Heinonen and Demetrius Albanes in 1994, the CARET randomized controlled trial by Gilbert S. Omenn in 1996, and the SELECT randomized controlled trial by Scott M. Lippman and Eric A. Klein in 2008. Similarly, these negative results of higher cancer incidence can be well explained by the same analysis. On the other hand, these clinical trials can verify from another perspective that the hypothesis that free radicals can cause cancer is correct. A natural antioxidant that makes the incidence of certain cancers higher can be used as a reference for how to design candidate drugs to intervene in these cancers in the future by analyzing its molecular structure.

[0087] Table 1. Randomized clinical trials of cancer intervention with natural antioxidants

[0088] 1994ATBC randomized controlled trial Beta-Carotene per Day 18% lung cancer incidence vs control group 1996CARET randomized controlled trial Beta-Carotene and Vitamin A Daily 28% lung cancer incidence vs control group SELECT 2008 randomized controlled trial Vitamin E 17% prostate cancer incidence vs control group

[0089] Free radical scavengers will definitely work as antioxidant drugs. The key is to effectively terminate free radicals and avoid the long-term propagation of free radicals, thereby greatly reducing the survival time of free radicals in the body. This logical analysis prompted the innovative design of the new aspirin mimetic of the present invention. Preclinical efficacy trials of the new drug have shown unprecedented results, such as reversal of cataracts in rat models, anti-inflammatory tests of in vitro cells, and excellent efficacy data when used compassionately in ALS patients.

[0090] The present invention is to treat ALS with a novel aspirin mimetic antioxidant via enteric-coated capsules and / or intravenous infusion. During the 6-month compassionate use of 6 ALS patients, oral administration of one capsule per day significantly slowed the progression of the disease and even slightly reversed ALS in some cases. Compared with edaravone, the new drug (DPL-002) has a significant efficacy of 67%. Given that there is currently no effective drug for the treatment of ALS, the current medical needs of this incurable motor neuron disease can be met by the new drug (DPL-002).

[0091] 1. Basic principles of free radical scavengers

[0092] The chemical activity of free radicals can usually be completely quenched by termination. The termination of free radicals is the key to the invention of new aspirin mimetics. A simple and effective way to terminate free radicals is to allow two free radicals to couple to form a stable coupled compound (Scheme 3). Disulfide bonds have the highest bond energy among the relevant functional groups considered, so thiols are the most powerful free radical scavengers.

[0093] As is well known, the antioxidant strength of thiophenol is much stronger than that of alkylthiols (such as cysteine, ALA, etc.). Therefore, thiophenol is the focus of the design of the new aspirin mimetic in the present invention. According to chemical kinetics, when the intramolecular disulfide bond forms a five-membered ring, the reaction rate of the dithiol-containing molecule to form a five-membered ring (intramolecular disulfide molecule) is much faster than the reaction rate of the formation of two molecules between molecules. In addition, the disulfide bond formed in the five-membered ring is much more stable than that formed in the open chain. Therefore, ALA is a better free radical scavenger than cysteine, because ALA can form intramolecular disulfide bonds in the five-membered ring, while cysteine ​​cannot. Similarly, the new aspirin mimetic in this project is a stronger free radical scavenger than ALA, because the thiophenol in the new aspirin mimetic is a stronger antioxidant than the alkylthiols in ALA, and they form a five-membered ring disulfide bond with a better bond angle.

[0094]

[0095] Scheme 3. Termination of free radicals by forming a coupling compound

[0096] Since thiophenol is a better antioxidant than alkylthiols, thioaspirin analogs can effectively terminate free radicals. The mechanism of effective termination of free radicals is shown in Scheme 4.

[0097]

[0098]

[0099] Scheme 4. New drug as an effective free radical scavenger

[0100] Therefore, the present invention applies the "drugability" of aspirin to a new antioxidant. In order to inherit the "drugability" of aspirin, the new antioxidant will retain the basic skeleton of the aspirin molecule, but replace the oxygen atoms in a pair of key functional groups with sulfur atoms. In the periodic table, sulfur and oxygen are in the same "oxygen family" and have similar properties. Replacing oxygen with sulfur will avoid any major "mutations" in the molecular structure, and the new antioxidant constructed in this way can still maintain the "drugability" of aspirin. As mentioned above, replacing oxygen with sulfur will make the new molecule the strongest free radical scavenger. In addition, the new molecule can participate in "disulfide bond exchange" (Scheme 5), which can reverse some misfolded proteins caused by abnormal disulfide bond cross-linking. Based on the Lewis Soft and Hard Acid-Base Theory (SHAB), replacing oxygen with sulfur will cause the new molecule to form a stronger chelate with heavy metals ( Fig.10 ). This is because sulfur is a softer base and has a higher affinity for heavy metal cations (soft acids) such as mercury, lead, cadmium, iron, and copper. Therefore, the new aspirin mimetic in this project can effectively remove heavy metals from the body. Chelation ability is delicate, as being too weak like aspirin or too strong like EDTA has been shown to be bad. Therefore, the active ingredient 2a of the new aspirin mimetic may be the ideal one.

[0101]

[0102] Solution 5: New drugs that participate in disulfide bond exchange can provide opportunities for correction of misfolded proteins

[0103] The antioxidant "drug" in the present invention should be its "reduced form". Disulfide bond compounds are inactive and no longer have the ability to quench free radicals. Thiols and thiophenols are "reduced forms" of antioxidants, but because they are easily oxidized by air, they are unstable in storage. Therefore, thiols and thiophenols must be chemically protected by other functional groups. At this point, the protected thiol compound is a prodrug that can release the "reduced form" of the active drug in the body, where the protecting group can be cut off by a biochemical reaction. The protecting group selected in the present invention also serves as a penetration enhancer, an efficacy enhancer, a selectivity enhancer, or a combination thereof. In addition, the protecting group must be safe and non-toxic.

[0104] 2. New aspirin mimetics

[0105] The novel aspirin mimetics of the present invention are summarized in several molecular structure models, such as Fig.11 Compounds DPL-001 and DPL-002 are the simplest forms of the novel aspirin mimetics of the present invention ( Fig.11 ). Compound 1a is a mimetic of salicylic acid and may be the active form of compound DPL-001 as an in vivo free radical scavenger. Compound 2a is also a mimetic of salicylic acid and may be the active form of compound DPL-002 as an in vivo free radical scavenger. Compounds 1b and 2b are known but are chemically prepared by different methods. The novel aspirin mimetic of the present invention is hydrolyzed in vivo to initially obtain compound 2a or 1a.

[0106] Schemes 4 and 6 propose a typical working mechanism of a new type of aspirin mimetic. Compound DPL-004 is delivered to target cells, such as neurons, where it is hydrolyzed to 2a. Compound 2a can effectively terminate two free radicals in vivo. Theoretically, in this regard, the dosage can be halved and the cost can also be halved. More importantly, it can effectively terminate free radicals and minimize the damage caused by primary and secondary free radicals.

[0107]

[0108] Scheme 6. Mechanism of action of new drugs

[0109] In summary, each new molecule of the present invention contains at least one pair of two functional groups. When oxidized, this configuration allows it to form a five-membered ring through a disulfide bond within the molecule. This pair of functional groups consists of a thiol (-SH) and a carbon disulfide (-CSSH) or a carbon sulfide (-COSH). Each pair of functional groups is covalently single-bonded to a carbon atom located on an aromatic ring or on a substituted aromatic ring. The aromatic ring can be a regular aromatic ring or an aromatic heterocycle, and any possible combination thereof.

[0110] Each pair of functional groups can be covalently bonded to other functional groups, including but not limited to acetyl (-Ac), amino acid residues, vitamin B residues, choline, dopamine, EDTA, carbohydrates, nucleobases, citric acid, succinic acid, 5-methoxytryptamine, heterocycles, etc. These functional groups can serve as protecting groups, penetration enhancers, efficacy enhancers, selectivity enhancers, or combinations thereof.

[0111] 3. Potential application of aspirin mimetic antioxidants in the intervention of ALS

[0112] Since free radicals may be one of the key causes of many major diseases, it is entirely possible to find an antioxidant drug that can scavenge free radicals and is effective against several diseases at the same time. It is admirable to find a class of antioxidant drugs with the same core structure and intervene in many major diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), diabetes, cancer, atherosclerosis, cardiovascular disease, kidney disease, hypertension, hyperlipidemia, rheumatoid arthritis, lupus, multiple sclerosis (MS), gout, inflammation, acne, pain, acne, stroke, depression, cataracts, glaucoma, macular degeneration and HIV, etc. The new antioxidants in the present invention are expected to show their talents in this field.

[0113] 3a. Amyotrophic lateral sclerosis

[0114] The cause of ALS is still unclear. Although there is no known cure, there are four FDA-approved drugs on the market. One of them is edaravone (Radicava), which was approved by the FDA in 2017 for the treatment of ALS and can slow the progression of ALS (early stage) by 33.3%. The working mechanism of edaravone is still unclear. Both the compounds of the present invention and edaravone are classified as antioxidants. In view of the molecular functions of the novel antioxidants explained above, the compounds of the present invention are better antioxidants than edaravone. Therefore, the compounds of the present invention are believed to be better than edaravone in treating amyotrophic lateral sclerosis.

[0115] Compared with edaravone, the compound of the present invention (DPL-002) showed an efficacy of 66.7% through compassionate use of patients. During the 6-month compassionate use of enteric-coated capsules (DPL-002), no adverse reactions were observed.

[0116] 4. Chemical Synthesis of the Novel Aspirin Mimics of the Present Invention

[0117] According to the procedure reported in JACS, dilithiation of thiophenol gave an ortho-directed lithiation intermediate. The dilithiation material reacted with a carbonyl sulfide and then acetylated (via acetic anhydride) to give compound 2 (DPL-002) (Scheme VII). Other target compounds can also be obtained when other corresponding acylating agents are used.

[0118]

[0119] Scheme 7. Synthesis of the novel antioxidant DPL-002

[0120] 5. ALS new drug efficacy trial

[0121] 5.1 Material preparation

[0122] The synthetic route of DPL-002 is shown in Scheme 7. The purity of DPL-002 exceeds 97%. Enteric-coated capsules are filled with corresponding doses of DPL-002; the enteric-coated capsule product does not contain other excipients.

[0123] DPL-001: 1H-NMR (400MHz, CD2Cl2) δ (ppm) 7.443 (s, 4H), 2.421 (d, J = 1.6Hz, 3H);

[0124] 13CNMR(100MHz,CD2Cl2)6193.850,134.538,129.371,129.113,129.083,128.210,127.512,127.239,29.989.

[0125] DPL-002: 1H-NMR (400MHz, CDCl3) δ (ppm) 8.0-8.15 (m, 1H), 7.55-7.65 (m, 2H), 7.45-7.55 (m, 1H), 2.46 (s, 3H); 13CNMR (100MHz, CDCl3) δ193.149,171,653,136.758,132.821,132.580,131.866,129.408,129.385,30.365.

[0126] DPL-003:1H-NMR(400MHz,DMSO-d6)δ(ppm)9.148(d,J=2.0Hz,1H),8.936(dd,J=1.6,5 .2Hz,1H),8.40-8.60(m,1H),7.70-7.90(m,1H),7.40-7.60(m,5H); 13CNMR(100MHz,DM SO-d6)δ186.322,150.983,146.113,144.557,135.446,133.420,130.750,128.625,1 28.208,128.109,124.802,124.240,123.739; MS(m / z)292(M+H)+,248(M+H-CS,100)+.

[0127] DPL-004: 1H-NMR (500MHz, CDCl3) δ (ppm) 9.179 (d, J=2.0Hz, 1H), 8.759 (dd, J= 2.0,5.0Hz,1H),8.07(dt,J=2.0,8.0Hz,1H),7.35-7.48(m,6H);13CNMR(125MH z, CDCl3)δ88.837,153.951,148.590,135.022(2),134.828,132.412,129.91 7(2),129.438(2),126.291,123.738; MS(m / z)274(MH)+,216(M+H-OCS,100)+.

[0128] 5.2 Compassionate Use Enteric Coated Capsules (DPL-002)

[0129] 1. Take one enteric-coated capsule daily with warm water 1 hour before breakfast.

[0130] 2. The capsules should be stored in a refrigerator (2-8°C) before use.

[0131] 3. Dose escalation process: April 18 to April 24, placebo for one week; April 25 to May 4, 10 mg per day for 10 consecutive days; May 5 to May 9, 20 mg per day for 5 consecutive days; May 10 to May 24, 40 mg per day for 15 consecutive days; May 25 to June 3, 50 mg per day for 10 consecutive days; June 4 to July 3, 100 mg per day; July 4 to July 31, 100 mg per day for 28 consecutive days; August 1 to September 4, 150 mg per day for 35 consecutive days; September 6 to September 30, 200 mg per day for 25 consecutive days, October 1 to October 20, 100 mg per day for 20 consecutive days.

[0132] 4. ALSFRS-R is scored monthly and the scores are recorded.

[0133] Table 1 ALSFRS-R scores

[0134]

[0135] 5.2 Data processing

[0136] ALSFRS-R score is an important indicator for measuring the condition of ALS patients. The change of this indicator before and after the use of the drug can reflect the efficacy. The method used in the 6-month compassionate use of enteric-coated capsules in the ALSFRS-R scoring process is the same as the method used in the 6-month Radicava (edaravone) clinical trial. The total change in the patient's ALSFRS-R score in Table 1 is (-10), and the average decrease is 6 points (-1.67). In the 6-month clinical trial of Radicava (edaravone), the average score was reduced to (-5.01). Using the same calculation method, the efficacy of the new drug capsule compared with edaravone is {5.01-1.67} / 5.01=66.7%. More than 25% is considered to have a significant efficacy. It is worth noting that according to the Wikipedia website (Edaravone-Wikipedia), Radicava's clinical trial for the treatment of ALS patients at all stages failed. In contrast, DPL-002's enteric-coated capsule product can intervene in various stages of ALS, as shown in Table 1.

[0137] General clinical observations: No adverse reactions were observed in all six ALS patients during the six months of continuous use of the enteric-coated capsules.

[0138] Conclusion: In terms of 6-month efficacy, the new compound DPL-002 is 66.7% more effective than edaravone in terms of ALSFRS-R score changes, significantly exceeding edaravone. In addition, DPL-002 capsules can intervene in all stages of ALS, rather than edaravone (oral or intravenous injection) which only targets early-stage ALS patients. In particular, patients 2 (early) and 4 (middle) had all ALSFRS-R score changes of zero, indicating no deterioration in 6 months; patient 5 (late) had an ALSFRS-R score change of 1, meaning a slight reversal of ALS progression. All these data indicate that DPL-002 is currently the best drug for ALS patients and can meet unmet medical needs.

[0139] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An aspirin mimetic antioxidant for treating amyotrophic lateral sclerosis, characterized in that: An active pharmaceutical ingredient selected from DPL-001, DPL-0012, DPL-003, DPL-004, or a pharmaceutically acceptable salt thereof; The molecular structure of DPL-001 is as follows: The molecular structure of DPL-002 is as follows: The molecular structure of DPL-003 is as follows: The molecular structure of DPL-004 is as follows:

2. An intravenous drip solution, characterized in that: The invention comprises the compound as claimed in claim 1, a pharmaceutically acceptable carrier and optionally one or more pharmaceutical excipients.

3. An intravenous drip solution according to claim 2, characterized in that: The pharmaceutically acceptable carrier is water, and the infusion solution further comprises a pharmaceutical excipient selected from sodium chloride, potassium chloride, sodium bisulfite and a combination thereof.

4. An intravenous drip solution according to claim 3, characterized in that: The dosage range of the active pharmaceutical ingredient is from 5 mg to 1000 mg per day, or from 0.1 mg / kg to 20 mg / kg body weight, or from 0.47 μmol / kg to 94 μmol / kg body weight.

5. An intravenous drip solution according to claim 4, characterized in that: The subjects were given once a day to 5 times a day.

6. A capsule, characterized in that: The invention comprises the compound as claimed in claim 1, a pharmaceutically acceptable carrier and optionally one or more pharmaceutical excipients.

7. A capsule according to claim 6, characterized in that: The pharmaceutically acceptable carrier is an enteric-coated capsule, wherein the enteric-coated capsule further comprises pharmaceutical excipients selected from starch powder, purified water, sodium chloride, potassium chloride, dicalcium phosphate, magnesium stearate, sodium carboxymethylcellulose and combinations thereof.

8. A capsule according to claim 7, characterized in that: The dosage of the active pharmaceutical ingredient is 10 mg to 2800 mg per day, or 0.2 mg / kg to 56 mg / kg body weight, or 0.94 micromol / kg to 263.2 micromol / kg body weight.

9. A capsule according to claim 8, characterized in that: The frequency of administration to the subjects ranged from once a day to 5 times a day.

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