Conjugate compounds comprising 9-cis-retinoic acid and monosaccharides

Abstract

The present disclosure provides a conjugated compound comprising 9-cis retinoic acid and a monosaccharide, and pharmaceutically acceptable salts thereof. The present invention also provides a pharmaceutical composition comprising the compound and a pharmaceutically acceptable carrier. The compound can be used for treating cancer or viral infection.

Description

Technical Field

[0001] This invention relates to a conjugated compound comprising 9-cis-retinoic acid and a monosaccharide, and a pharmaceutically acceptable salt thereof. This invention also relates to a pharmaceutical composition comprising said compound and a pharmaceutically acceptable carrier. Background Technology

[0002] Retinoic acid comprises all-trans-retinoic acid (ATRA, also known as vitamin A acid or tretinoin) and 9-cis-retinoic acid. Due to its specific effects on cell proliferation, differentiation, and apoptosis, and its low toxicity, it has become a promising compound for the treatment of various cancers. Biochemists have discovered retinoic acid receptors in the nucleus of human cells, which do not mutate in cancer cells.

[0003] Several clinical trials are currently underway that incorporate ATRA as part of combination therapies. For example, ATRA combined with different interferons (IFNs) has been shown to enhance anti-cancer effects, including growth inhibition and cell death in tumor cell lines. However, to fully realize the therapeutic potential of retinoic acid, many studies emphasize the need to better understand the mechanisms by which retinoic acid signaling and gene expression are blocked in cancers such as acute myeloid leukemia (AML). These studies demonstrate that combination therapies targeting multiple gene silencing mechanisms are often the most effective strategy for reactivating ATRA-sensitive gene expression and AML cell differentiation, as well as mediating the general anti-cancer activity of ATRA.

[0004] The identification of protein classes that control gene expression via histones and DNA modifications has driven the development of novel therapeutic agents that alter chromatin structure, known as epigenetic drugs. However, these epigenetic drugs have been shown to be only partially effective against various cancers when used alone. This limitation underscores the need to explore alternative approaches that can complement or enhance cancer therapies. Summary of the Invention

[0005] This invention relates to conjugated compounds comprising 9-cis-retinoic acid and a monosaccharide. The 9-cis-retinoic acid in the conjugated compound provides therapeutic activity, and the monosaccharide in the conjugated compound increases the water solubility of the compound and significantly reduces the effective dose of 9-cis-retinoic acid in the compound for treating diseases.

[0006] In one embodiment, the compound is represented by formula (I):

[0007] (I)

[0008] Among them, R1 R is a monosaccharide, whether substituted or unsubstituted. 2 It is a substituted or unsubstituted aliphatic group having 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms, and more preferably 1 to 6 carbon atoms, and n is 0 or 1. Attached Figure Description

[0009] Figure 1 The 1H NMR spectrum of galactose-9-cis-retinoic acid was described.

[0010] Figure 2 A graph illustrating cell survival rates according to an embodiment of the present invention is provided, showing the cytotoxic effect of galactose-9-cis-retinoic acid on the A549 cancer cell line.

[0011] Figure 3 A graph illustrating cell survival rates according to an embodiment of the present invention is provided, showing the cytotoxic effect of galactose-9-cis-retinoic acid on the H460 cancer cell line.

[0012] Figure 4 A graph illustrating cell survival rates according to an embodiment of the present invention is provided, showing the cytotoxic effect of galactose-9-cis-retinoic acid on the Huh-7 cancer cell line.

[0013] Figure 5 A graph illustrating cell survival rates according to an embodiment of the present invention is provided, showing the cytotoxic effect of galactose-9-cis-retinoic acid on the AsPC-1 cancer cell line.

[0014] Figure 6 A graph illustrating cell survival rates according to an embodiment of the present invention is provided, showing the cytotoxic effect of galactose-9-cis-retinoic acid on the MDA-MB-231 cancer cell line.

[0015] Figure 7 A graph illustrating cell survival rates according to an embodiment of the present invention is provided, showing the cytotoxic effect of galactose-9-cis-retinoic acid on the SKOV-3 cancer cell line. Detailed Implementation

[0016] Conjugated compounds

[0017] This invention relates to conjugated compounds comprising 9-cis-retinoic acid and a monosaccharide. The 9-cis-retinoic acid in the conjugated compound provides therapeutic activity, and the monosaccharide in the conjugated compound increases the water solubility of the compound and significantly reduces the effective dose of 9-cis-retinoic acid in the compound for treating diseases.

[0018] In one embodiment, the compound is represented by formula (I):

[0019] (I)

[0020] Among them, R 1 R is a monosaccharide, whether substituted or unsubstituted. 2 It is a substituted or unsubstituted aliphatic group having 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms, and more preferably 1 to 6 carbon atoms, and n is 0 or 1.

[0021] Monosaccharides (also known as simple sugars) are the simplest form of sugar and the most basic building blocks (monomers) of all carbohydrates. Chemically, monosaccharides contain substances with the chemical formula H-[CHOH]. n -CHO polyhydroxy aldehydes or those with the chemical formula H-[CHOH] m -CO-[CHOH] n -H polyhydroxy ketones with three or more carbon atoms.

[0022] In some embodiments, the monosaccharides include, but are not limited to: aldoses such as galactose, glucose, mannose, glyceraldehyde, erythrose, ribose, arabinose, xylose, idose, gulose, talose, allose, and altrose; ketoses such as fructose, erythrulose, ribulose, xylulose, pscicose, sorbose, and tagatose; deoxysaccharides such as deoxyribose, 2-deoxyglucose, fucose, rhamnose, and quinovose; and aminosaccharides such as glucosamine, galactosamine, and mannose. Preferred monosaccharides of the present invention include galactose, glucose, mannose, fructose, ribose, deoxyribose, and fucose.

[0023] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof may be prepared in dimethyl sulfoxide (DMSO), ethanol, buffer solution, saline, or water for administration, the dosage and frequency of which may vary depending on factors such as the severity of the disease to be treated (e.g., cancer) and the weight, age, physical condition, and response of the subject to be treated. The daily dose of the above therapeutic agent may be administered once or multiple times.

[0024] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered orally, intravenously, intramuscularly, subcutaneously, intraperitoneally, intranasally, as an aerosol, or topically.

[0025] Pharmaceutical Composition

[0026] The present invention provides a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers and compounds of formula (I).

[0027] In one embodiment, the compound is incorporated into any acceptable carrier, comprising cream, gel, lotion, or other type of suspension, said carrier stabilizing the compound and delivering it to the affected site via local application. In another embodiment, the pharmaceutical composition may be in dosage forms such as lozenges, capsules, granules, fine particles, powders, syrups, suppositories, injections, patches, etc. The above pharmaceutical compositions can be prepared using conventional methods.

[0028] Pharmaceutically acceptable carriers, which are inactive ingredients, can be selected by those skilled in the art using conventional standards. Pharmaceutically acceptable carriers include, but are not limited to, non-aqueous solutions, suspensions, emulsions, microemulsions, microcellular solutions, gels, and ointments. Pharmaceutically acceptable carriers may also contain, but are not limited to, the following components: saline and aqueous electrolyte solutions; ionic and nonionic permeabilizers, such as sodium chloride, potassium chloride, glycerol, and glucose; pH adjusters and buffers, such as hydroxides, phosphates, citrates, acetates, and borates; triethanolamine; and antioxidants, such as bisulfite, sulfite, metabisulfite, thiosulfite, ascorbic acid, acetylcysteine, cysteine, glutathione, butylated hydroxyanisole, butylated hydroxytoluene, tocopherol, and ascorbyl palmitate. Salts, acids, and / or bases of palmitate; surfactants such as lecithin and phospholipids, including but not limited to phosphatidylcholine, phosphatidylethanolamine, and phosphatidyl... Inositiol); poloxamer and poloxamine; polysorbates, such as polysorbate 80, polysorbate 60 and polysorbate 20; polyethers, such as polyethylene glycol and polypropylene glycol; polyethylene, such as polyvinyl alcohol and polyvinylpyrrolidone; cellulose derivatives, such as methylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, carboxymethylcellulose and hydroxypropylmethylcellulose and their salts; petroleum derivatives, such as mineral oil and white petrolatum; fats, such as lanolin, peanut oil, palm oil and soybean oil; mono-, di- and triglycerides; acrylic polymers, such as carboxymethyl polymethyl gel and hydrophobically modified cross-linked acrylic copolymers; polysaccharides, such as dextran and glycosaminoglycans, such as sodium hyaluronate. Other pharmaceutically acceptable carriers include xanthan gum, carrageenan, Avicel RC-591 (a combination of microcrystalline cellulose and polyethylene glycol).Alternatively, the active compound may be soluble or suspended in pharmaceutically acceptable lipid formulations, such as those described by Kalepu et al. (Acta Pharmaceutica Sinica B, 3: 361-372, 2013), for example, vegetable oils, coconut oil, castor oil, etc.

[0029] Such pharmaceutically acceptable carriers may use familiar preservatives to prevent bacterial contamination, including but not limited to: benzalkonium chloride, ethylenediaminetetraacetic acid and its salts, benzethonium chloride, chlorhexidine, chlorobutanol, methylparaben, thimerosal and phenethyl alcohol, or may be formulated into non-preservative preparations for single or multiple use.

[0030] For example, the tablet or capsule formulation of the compound may contain other non-biologically active excipients that do not react with the compound. The excipients for the tablets or capsules may include fillers, binders, lubricants and glidants, disintegrants, wetting agents, and release modifiers. Binders promote the adhesion of the formulation particles and are crucial for tablet formulations. Examples of excipients for tablets or capsules include, but are not limited to: carboxymethyl cellulose, cellulose, ethyl cellulose, hydroxypropyl methylcellulose, methyl cellulose, karaya gum, starch, tragacanthum, gelatin, magnesium stearate, titanium dioxide, polyacrylic acid, and polyvinylpyrrolidone. For example, tablet formulations may contain inactive ingredients such as colloidal silica, crospovidone, hydroxypropyl methylcellulose, magnesium stearate, microcrystalline cellulose, polyethylene glycol, sodium starch glycolate, and / or titanium dioxide. Capsule formulations may contain inactive ingredients such as gelatin, magnesium stearate, and / or titanium dioxide.

[0031] For example, the patch formulation of the compound may contain inactive ingredients such as 1,3-butanediol, aluminum hydroxyacetate, disodium edetate, D-sorbitol, gelatin, kaolin, methylparaben, polysorbate 80, polyvinylpyrrolidone, propylene glycol, propylparaben, sodium carboxymethyl cellulose, sodium polyacrylate, tartaric acid, titanium dioxide, and purified water. The patch formulation may also contain skin penetration enhancers such as lactate esters or diethylene glycol monoethyl ether.

[0032] Topical formulations containing the aforementioned compounds may be in the form of gels, creams, lotions, liquids, emulsions, ointments, sprays, solutions, and suspensions. Inactive ingredients in topical formulations may include, but are not limited to: (emollients / penetration enhancers), diethylene glycol monoethyl ether (emollients / penetration enhancers), DMSO (solution enhancers), polysiloxane elastomers (rheology modifiers / texture modifiers), caprylic / capric triglycerides (emollients), octisalate (emollients / UV filters), polysiloxane liquids (emollients / diluents), squalene (emollients), sunflower oil (emollients), and silica (thickeners).

[0033] How to use

[0034] The claimed compound can be used to treat cancers such as pancreatic cancer, lung cancer, breast cancer, liver cancer, and ovarian cancer. The method includes the step of administering a therapeutically effective amount of the claimed compound to a cancer patient. As used herein, "effective amount" means an amount that effectively treats the disease by improving the pathological condition or alleviating disease symptoms.

[0035] The claimed compound can also be used to treat viral infections, such as respiratory syncytial virus (RSV). The method includes administering a therapeutically effective amount of the claimed compound to a subject in need, wherein the effective amount is sufficient to inhibit viral replication, reduce viral load, or alleviate infection-related symptoms. As used herein, "effective amount" means an amount that effectively reduces the pathological condition caused by a viral infection or improves the clinical symptoms of an infected subject.

[0036] The pharmaceutical compositions of the present invention can be administered via systemic or regional application. Regional application includes local application and inhalation. Systemic application includes oral, non-enteric (such as intravenous, intramuscular, subcutaneous, or rectal) and other systemic routes of administration. In systemic application, the active compound first reaches the plasma and then distributes to the target tissue. The preferred route of administration of the present invention is intravenous administration.

[0037] The administration of the composition may vary depending on the degree of harm and the individual response of each patient. For systemic administration, the plasma concentration of the delivered active compound may vary; however, it is typically 1 × 10⁻⁶. -10 Up to 1 × 10 -4 moles per liter, and preferably 1 × 10⁻⁶. -8 Up to 1 × 10 -5 Moles per liter.

[0038] In one embodiment, the pharmaceutical composition is administered to the subject intravenously. The dose administered by intravenous bolus or infusion is typically from 0.01 to 100 mg / kg / day, and preferably from 0.03 to 30 mg / kg / day.

[0039] In one embodiment, the pharmaceutical composition is administered orally to the subject.

[0040] Those skilled in the art will understand that various types of delivery mechanisms are also applicable to this invention.

[0041] This method can be used to treat mammalian subjects, such as humans, horses, and dogs. This invention is particularly suitable for treating humans.

[0042] It should be understood that the above presents preferred embodiments of the present invention, and modifications may be made thereto without departing from the scope of the invention as described in the claims.

[0043] This invention provides at least the following advantages:

[0044] 1. The claimed compounds and their pharmaceutical compositions can enhance the ability to inhibit cancer cells, thereby effectively treating a variety of cancers, such as lung cancer, ovarian cancer, breast cancer, pancreatic cancer, and liver cancer.

[0045] 2. The claimed compounds and their pharmaceutical compositions have antiviral activity, particularly against respiratory syncytial virus (RSV).

[0046] 3. The proposed compound has better water solubility, thereby reducing the effective dose of retinoic acid and minimizing side effects. This improvement increases the efficiency and concentration of retinoic acid as it crosses cell membranes and enters cells for therapeutic purposes.

[0047] Example

[0048] The present invention will be further illustrated by the following examples. However, it should be understood that the following examples are for illustrative purposes only and should not be construed as limiting the invention in practice.

[0049] Example 1: Synthesis of galactose-9-cis-retinoic acid

[0050] Materials and Methods

[0051] Galactose-9-cis-retinoic acid, i.e., a conjugated compound of galactose linked to 9-cis-retinoic acid, was synthesized via the following procedure I:

[0052]

[0053] Process I

[0054] Step 1: Synthesis of Compound 2: 2,5-di-side-oxypyrrolidin-1-yl(2E,4E,6Z,8E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohexyl-1-en-1-yl)non-2,4,6,8-tetraenoic acid ester

[0055]

[0056] HOSu (29 mg, 0.25 mmol), DCC (69 mg, 0.34 mmol), and 4-DMAP (21 mg, 0.17 mmol) were added to anhydrous DMF (2 mL) containing 9-cis-retinoic acid (50 mg, 0.17 mmol). The reaction mixture was stirred at 25 °C for 16 hours. The reaction mixture was treated with H₂O and extracted with EtOAc. The combined organic phases were washed twice with brine, dried over Na₂SO₄, and concentrated under vacuum to give the title compound (60 mg, 0.12 mmol, 73% yield) as a yellow solid, which could be used directly without further purification.

[0057] Step 2: Synthesis of Compound 4: (2R,3S,4S,5R,6R)-2-(acetoxymethyl)-6-bromotetrahydro-2H-piperan-3,4,5-triacetic acid triester

[0058]

[0059] A 33% AcOH solution containing HBr (8 mL) and acetic anhydride (1.5 mL) were added to a DCM solution (30 mL) containing (2S,3R,4S,5S,6R)-6-(acetoxymethyl)tetrahydro-2H-piperan-2,3,4,5-tetraacetic acid tetraester (5 g, 12.8 mmol). The reaction mixture was stirred at 25 °C for 6 hours. The reaction mixture was poured into ice water and extracted with DCM. The combined organic layers were dried over MgSO4, filtered, and concentrated. The residue was purified by rapid chromatography (elution buffer: EtOAc dissolved in PE = 0 to 60%) to give the title compound (5.5 g, 12 mmol, 93% yield) as a colorless oil.

[0060] 1H NMR (400 MHz, CDCl3) δ 6.70 (d, J = 4.0 Hz, 1H), 5.52 (d, J = 3.2Hz, 1H), 5.41 (dd, J = 10.8, 3.2 Hz, 1H), 5.05 (dd, J = 10.4, 4.0 Hz, 1H), 4.49 (t, J = 6.4 Hz, 1H), 4.14 – 4.07 (m, 2H), 2.15 (s, 3H), 2.12 (s, 3H), 2.06 (s, 3H), 2.01 (s, 3H).

[0061] Step 3: Synthesis of Compound 5: Methyl [(3S,4S,6R)-3,4,5-tris(acetoxy)-6-(3-bromopropoxy)oxacyclohexyl-2-yl]acetate

[0062]

[0063] 3-Bromoprop-1-ol (1.7 g, 12.2 mmol) was added to a 50 mL solution of dichloromethane containing compound 4 (5 g, 12.2 mmol) and powdered 3A MS (5 g). TLC examination of the mixture showed no starting material. AgOTf (6.27 g, 24.4 mmol) was added to the mixture at 0 °C. The mixture was stirred at 25 °C for 16 h. The residue was diluted with water and extracted with DCM. The combined organic layers were dried over Na₂SO₄, filtered, and concentrated. The residue was purified by rapid chromatography (elution buffer: EtOAc dissolved in PE = 0 to 80%) to give the title compound (1.5 g, 2.9 mmol, 24% yield) as a yellow oil.

[0064] 1H NMR (400 MHz, CDCl3) δ 5.40 (d, J = 3.2 Hz, 1H), 5.20 (dd, J =10.4, 8.0 Hz, 1H), 5.03 (dd, J = 10.4, 3.2 Hz, 1H), 4.48 (d, J = 8.0 Hz, 1H),4.22 – 4.10 (m, 4H), 4.05 – 3.98 (m, 1H), 3.92 (t, J = 6.4 Hz, 1H), 3.72 –3.66 (m, 1H), 3.50 – 3.46 (m, 2H), 2.15 (s, 3H), 2.09 (s, 3H), 2.06 (s, 3H), 1.99 (s, 3H).

[0065] Step 4: Synthesis of Compound 6: Methyl [(3S,4S,6R)-3,4,5-tris(acetoxy)-6-(3-azidopropoxy)oxacyclohexyl-2-yl]acetate

[0066]

[0067] NaN3 (0.42 g, 6.4 mmol) was added to anhydrous DMF (15 mL) containing compound 5 (1.5 g, 3.2 mmol), and the mixture was stirred at 100 °C for 16 h. The reaction mixture was treated with H2O and extracted with EtOAc. The combined organic phases were washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by rapid chromatography (elution: EtOAc dissolved in PE = 0 to 90%) to give the title compound (0.64 g, 1.3 mmol, 41% yield) as a yellow oil.

[0068] 1H NMR (400 MHz, CDCl3) δ 5.39 (d, J = 2.8 Hz, 1H), 5.20 (dd, J =10.4, 8.0 Hz, 1H), 5.02 (dd, J = 10.4, 3.2 Hz, 1H), 4.47 (d, J = 8.0 Hz, 1H),4.22 – 4.13 (m, 2H), 4.02 – 3.88 (m, 2H), 3.64 – 3.58 (m, 1H), 3.41 – 3.34(m, 2H), 2.16 (s, 3H), 2.07 (s, 3H), 2.05 (s, 3H), 1.99 (s, 3H), 1.91 – 1.81(m, 2H).

[0069] Step 5: Synthesis of Compound 7: (2R,4S,5R)-2-(3-azidopropoxy)-6-(hydroxymethyl)oxacyclohexane-3,4,5-triol

[0070]

[0071] NaOMe (401 mg, 7.4 mmol) was added to a solution of MeOH (8 mL) and DCM (8 mL) containing compound 6 (640 mg, 1.48 mmol). The reaction mixture was stirred at 25 °C for 16 hours. The reaction mixture was then analyzed using Dowex... ®The pH of the reaction mixture was acidified to 7 using 50W x 8. The mixture was filtered and concentrated under vacuum. The residue was purified by rapid chromatography (elution buffer: MeOH dissolved in DCM = 0 to 20%) to give the title compound (350 mg, 1.19 mmol, 80% yield) as a yellow oil.

[0072] 1H NMR (400 MHz, CD3OD) δ 4.21 (d, J = 7.2 Hz, 1H), 3.99 – 3.93 (m,1H), 3.83 (d, J = 3.2 Hz, 1H), 3.75 – 3.71 (m, 2H), 3.66 – 3.61 (m, 1H), 3.53–3.43 (m, 5H), 1.90–1.83 (m, 2H).

[0073] Step 6: Synthesis of Compound 8: (2R,4S,5R)-2-(3-aminopropoxy)-6-(hydroxymethyl)oxacyclohexane-3,4,5-triol

[0074]

[0075] Pd(OH)₂ / C (94 mg, 20 wt%) was added to a 5 mL solution of MeOH containing compound 7 (350 mg, 1.33 mmol). The mixture was stirred at 25 °C for 16 hours. The reaction mixture was filtered through a diatomaceous earth pad and washed with MeOH. The filtrate was concentrated under vacuum to give the title compound (250 mg, 0.84 mmol, 64% yield) as a colorless oil, which was used in the next step without further purification.

[0076] 1H NMR (400 MHz, CD3OD) δ 4.21 (d, J = 7.2 Hz, 1H), 4.03 – 3.94 (m,1H), 3.82 (d, J = 3.2 Hz, 1H), 3.76 – 3.70 (m, 2H), 3.67 – 3.63 (m, 1H), 3.52–3.43 (m, 3H), 2.81–2.76 (m, 2H), 1.80–1.74 (m, 2H).

[0077] Step 7: Synthesis of target compound Ia: (2E,4E,6Z,8E)-3,7-dimethyl-N-(3-{[3,4,5-trihydroxy-6-(hydroxymethyl)oxacyclohexyl-2-yl]oxy}propyl)-9-(2,6,6-trimethylcyclohexyl-1-en-1-yl)non-2,4,6,8-tetraenoamide

[0078]

[0079] 2,5-di(2E,4E,6Z,8E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohexyl-1-en-1-yl)non-2,4,6,8-tetraenoic acid ester (60 mg, 0.15 mmol) and triethylamine (52 mg, 0.51 mmol) were added to anhydrous DMF (3 mL) containing compound 8 (40 mg, 0.17 mmol). The reaction mixture was stirred in an ice bath for 3 hours. The mixture was treated with H2O and extracted with EtOAc. The residue was purified by rapid chromatography (elution: MeOH in DCM = 0 to 20%) to give the title compound (20 mg, 0.04 mmol, 22% yield) as a yellow solid.

[0080] MS (ESI): C 29 H 45 The theoretical mass of NO7 is 519.32, and the observed m / z value is 542.35 [M+Na]. + .

[0081] ( Figure 1)1H NMR (400 MHz, CD3OD-d4) δ 7.06 – 6.99 (m, 1H), 6.68 (d, J =16.0 Hz, 1H), 6.29 – 6.23 (m, 2H), 6.08 (d, J = 11.6 Hz, 1H), 5.84 (s, 1H),4.22 (d, J = 7.6 Hz, 1H), 3.96 – 3.90 (m, 1H), 3.82 (d, J = 3.2 Hz, 1H), 3.78– 3.70 (m, 2H), 3.66 – 3.60 (m, 1H), 3.54 – 3.44 (m, 3H), 3.41 – 3.32 (m,2H), 2.27 (s, 3H), 2.06 (t, J = 6.4 Hz, 2H), 1.98 (s, 3H), 1.84 – 1.78 (m,2H), 1.74 (s, 3H), 1.69 – 1.62 (m, 2H), 1.53 – 1.48 (m, 2H), 1.04 (s, 6H); 1HNMR (400 MHz, DMSO-d6) δ 7.89 (t, J = 5.2 Hz, 1H), 7.01 – 6.90 (m, 1H), 6.70(d, J = 16.0 Hz, 1H), 6.26 – 6.20 (m, 2H), 6.12 (d, J = 11.2 Hz, 1H), 5.82(s, 1H), 4.86 (d, J = 2.8 Hz, 1H), 4.69 (d, J = 3.6 Hz, 1H), 4.58 (t, J = 5.6Hz, 1H), 4.35 (d, J = 4.4 Hz, 1H), 4.07 (d, J = 6.0 Hz, 1H), 3.79 – 3.73 (m,1H), 3.62 (s, 1H), 3.55 – 3.45 (m, 3H), 3.28 – 3.23 (m, 3H), 3.21 – 3.11 (m,2H), 2.24 (s, 3H), 2.05 – 2,00 (m, 2H), 1.95 (s, 3H), 1.72 (s, 3H), 1.69 –1.64 (m, 2H), 1.63 – 1.55 (m, 2H), 1.47 – 1.42 (m, 2H), 1.02 (s, 6H)。

[0082] Monosaccharides share several structural similarities, containing multiple hydroxyl (-OH) groups attached to their carbon backbone. The main difference lies in the number and orientation of these hydroxyl groups. Therefore, the synthetic flow chart for other monosaccharides, 9-cis-retinoic acid, follows a method similar to that outlined in Example 1.

[0083] Example 2: Evaluation of the in vitro cytotoxic effects of galactose-9-cis-retinoic acid on different cancer cell lines

[0084] In this example, seven cell lines were used: A549, NCI-H460, Huh-7-Ab, AsPC-1-Ab, MDA-MB-231-Ab, and a cancer cell line. Each cell line was cultured in a specified growth medium containing 10% FBS in a humidified incubator at 37°C containing 5% CO2.

[0085] The day before processing, cells in the logarithmic growth phase were collected, counted, and divided into 1 × 10⁻⁶ cells. 4 Cells were seeded at a density of 100 μL / well in 96-well plates. After incubation overnight, the culture medium was gently removed from each well, and then fresh culture medium (200 μL / well) was added.

[0086] The test article (TA) was a novel retinoid compound (referred to as the "novel compound" in this example), which was freshly prepared as a 48 mM stock solution in 100% DMSO on the day of treatment. The novel compound was serially diluted twofold using 100% DMSO to obtain concentrations ranging from 0.375 mM to 48 mM. Cells were treated by adding 2.02 μL of the specified concentration of the novel compound to each well to obtain final concentrations ranging from 3.75 μM to 480 μM, while maintaining 1% DMSO in all wells (including a DMSO control). All components were gently mixed and incubated for 24 hours.

[0087] On the day of cell viability measurement, the culture medium in each well was replaced with freshly prepared Alamarblue dye (10% v / v), and the cells were incubated at 37°C for 2 to 3 hours. Spectrophotometric absorbance was recorded at wavelengths of 570 nm and 600 nm.

[0088] Calculate the percentage of cell viability using the following formula:

[0089] Survival rate % = (TA of [A]) 570 / A 600 [Ratio] / DMSO of [A] 570 / A 600 Ratio])*100%

[0090] Where TA = item to be tested

[0091] A 570 absorbance at 570 nm

[0092] A 600 absorbance at 600 nm

[0093] Results: See Figures 2 to 7 Studies have shown that novel compounds can significantly reduce cell survival rates in all six cancer cell lines, indicating that novel retinoic acid compounds, namely galactose-modified 9-cis-retinoic acid, may be potential anticancer drugs for the effective treatment of various cancers, such as lung cancer, ovarian cancer, breast cancer, pancreatic cancer, and liver cancer. Table 1 shows the IC50 values. 50 .

[0094] Table 1

[0095]

[0096] Example 3: Evaluation of the in vitro cytotoxic effects of galactose-9-cis-retinoic acid on antiviral cytotoxicity.

[0097] The antiviral efficacy of galactose-9-cis-retinoic acid against respiratory syncytial virus (RSV) strain B1 was tested in HEp2 cells.

[0098] In short, the antiviral activity of galactose-9-cis-retinoic acid was compared. A stock solution of galactose-9-cis-retinoic acid at a concentration of 96 mM was prepared using DMSO (Sigma, D2650).

[0099] First, in order to perform viral infection, HEp2 cells were cultured at a density of 1.5 × 10⁻⁶. 5Cells were seeded at a density of 10 cells / well in 12-well plates. Cells were then cultured for one day in an incubator maintained at 37°C and 5% CO2. After washing with Dulbecco's Phosphate-Buffered Saline (DPBS), RSV B1 with a multiplicity of infection (MOI) of 0.1 and 50 µM or 100 µM galactose-9-cis-retinoic acid (GNA) or DMSO (vehicle) were added to the cells. Cells were cultured for an additional 1 hour in an incubator at 37°C and 5% CO2, followed by washing with DPBS. Subsequently, 50 µM or 100 µM galactose-9-cis-retinoic acid (GNA) or DMSO (vehicle) was added to 1 mL of 10% fetal bovine serum (FBS) / Dulbecco's Modified Eagle Medium (DMEM). Cells were cultured at 37°C in a 5% CO2 incubator for 24, 48, and 72 hours. At the end of the experiment, the culture medium was removed, and the cells were washed again with DPBS. Total RNA from the cultured cells was collected by adding 500 μL of RNAzol, and the lysate was collected. The collected lysate was stored at -80°C.

[0100] Subsequently, RSV content in each group of cells was assessed. First, RNA isolation was performed by thawing cell lysates containing total RNA at room temperature and adding 200 μL of diethyl pyrocarbonate (DEPC)-H2O. The mixture was vortexed and allowed to stand for 15 minutes. Then, the mixture was centrifuged at 12,000 ×g for 15 minutes at 4°C. Next, 200 μL of 75% EtOH was added to a new microcentrifuge tube (Eppendorf), followed by 500 μL of supernatant. The mixture was allowed to stand for 10 minutes, then centrifuged at 12,000 ×g for 8 minutes at 4°C. The supernatant was removed, and the precipitate was washed twice with 500 μL of 75% EtOH. Subsequently, RNA was dissolved in 12 μL of DEPC-H2O, and RNA quantification was performed. Unused RNA was stored at -80°C.

[0101] The isolated RNA was subjected to reverse transcription (RT). First, 4 μg of RNA was diluted to 8 μL with DEPC-H2O, and 1 μL of 10× RT reaction buffer and 1 μL of DNase I were added. The reaction mixture was incubated at 37°C for 30 minutes. Subsequently, 1 μL of 50 mM EDTA was added, and the mixture was incubated at 65°C for another 10 minutes. Then, 1 μL of random hexamer, dNTPs, and DEPC-H2O were added to the reaction mixture, and the mixture was incubated at 65°C for 5 minutes. Next, 4 μL of 5× first-strand buffer, 1 μL of dithiothreitol (DTT), and GScript RTase were added. The reaction mixture was incubated at 50°C for 50 minutes, followed by incubation at 70°C for 15 minutes. The reverse transcription product was stored at -80°C until further analysis. Finally, following standard procedures and using the SYBR system with a QuantStudio 6 Flex real-time PCR system, the RSV content in each treated cell group was assessed by qPCR, with actin used as a control. The qPCR program was as follows: 95℃ for 10 seconds, followed by 60℃ for 20 seconds and a 72℃ extension for 1 second, repeated for 45 cycles. The primer sequences used for RSV and actin are shown in Table 2 below.

[0102] Table 2

[0103]

[0104] qPCR results using actin and RSV N primers provided the relative RSV RNA content in different groups of cultured cells receiving different treatments. As shown in Table 3 below, cells in group 2 treated with 50 µM galactose-9-cis-retinoic acid showed lower RSV RNA content compared to the carrier group. Treatment with 50 µM galactose-9-cis-retinoic acid effectively inhibited RSV replication, suppressing 98.4% of RSV immediately after treatment, 99.0% after 48 hours, and 94.7% after 72 hours. When the compound concentration was increased to 100 µM (group 3), galactose-9-cis-retinoic acid showed a significant viral inhibition rate (greater than 99%).

[0105] Table 3

[0106]

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound is represented by formula (I): （I） Among them, R 1 R is a monosaccharide. 2 It is a substituted or unsubstituted aliphatic group having 1 to 20 carbon atoms, and n is 0 or 1.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The monosaccharides are selected from the group consisting of: galactose, glucose, mannose, glyceraldehyde, fucose, ribose, arabinose, xylose, idole, gulose, talose, allose, azoose, fructose, fucoulose, ribulose, xylulose, pseudofructose, sorbose, tagatose, deoxyribose, 2-deoxyglucose, fucose, rhamnose, quinoose, glucosamine, galactosamine, and mannose.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The monosaccharide is galactose, glucose, mannose, fructose, ribose, deoxyribose, or fucose.

4. A pharmaceutical composition comprising the compound according to claim 1, 2 or 3 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

5. A method of treating a subject with cancer, comprising administering to the subject a therapeutically effective amount of the compound according to claim 1, 2 or 3 or a pharmaceutically acceptable salt thereof.

6. The method according to claim 5, characterized in that, The cancer in question is lung cancer, liver cancer, pancreatic cancer, breast cancer, or ovarian cancer.

7. A method of treating a subject with a viral infection, comprising administering to the subject in need a therapeutically effective amount of the compound of claim 1, 2 or 3 or a pharmaceutically acceptable salt thereof.

8. The method according to claim 7, characterized in that, The virus in question is respiratory syncytial virus (RSV).

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