Preparation of novel triterpene alcohol derivatives with improved bioavailability for the treatment of cancer, inflammation and pain
By polar or charged modification of tetracyclic terpene alcohol compounds, the problems of their water solubility and low bioavailability are solved, and more efficient bioavailability and therapeutic effects are achieved, especially in cancer treatment, which shows significant inhibitory activity.
Patent Information
- Application Number
- JP2024547095
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2022-02-07
- Filing Date
- 2023-02-07
- Publication Date
- 2025-02-06
AI Technical Summary
Existing natural compounds such as tetracyclic terpene alcohols have problems with low bioavailability in terms of water solubility and permeability, resulting in high production costs and unstable effects.
By introducing polar or charged partial structures, tetracyclic terpene alcohols are modified to form modified compounds such as sulfate, phosphate, amino acid complex, PEG polymer, cyclodextrin complex and diacid derivatives to improve their solubility and bioavailability in water.
The water solubility and bioavailability of the compounds are significantly improved, showing potential effects on cancer, inflammation and pain treatment, especially at low doses to specific cancer cells.
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Abstract
Description
[Technical field]
[0001] The present invention relates to the pharmaceutical use of tetracyclic terpene 3-ols, typically having polar and / or charged substructures, such as lanosta-8,24-dien-3-ol, as anti-inflammatory, anti-cancer and analgesic agents via inhibition of the unregulated activation of serine-threonine protein kinases, particularly PKC. [Background technology]
[0002] Natural compounds such as tetracyclic triterpenes have attracted much attention due to their anti-cancer and anti-inflammatory activities. However, pharmacokinetic studies have shown poor oral bioavailability associated with their poor solubility and permeability in aqueous media. Their formulations are highly hit-to-miss and highly dependent on the excipients and manufacturing methods used, resulting in high overall manufacturing costs. It is essential to optimize the structure of the parent compound to increase water solubility. Summary of the Invention
[0003] Triterpene alcohols are practically insoluble in water. We have explored the introduction of polar or charged substructures to selected triterpene alcohol groups. Such derivatives with polar, negatively or positively charged groups have the ability to mix with water and lipids, similar to any detergent molecule. Preferred modification groups are selected from those with transient stability, which can be influenced by the tissue environment and / or local enzymatic activity. The proposed modifications R strive to keep the core triterpene structure intact, as shown below.
[0004] [ka] wherein R--a preferred polar or charged moiety; and the R1, R2, R3, R4, R5, R6, R7, R8 moieties are hydrogen, hydroxyl, methyl group, hydroxymethyl, carboxyl, or combinations thereof.
[0005] 1. Triterpene sulfates. Triterpene sulfates exist as metabolic products in some species. Sulfate esters of steroids are known to be more soluble in water than the steroids themselves. Triterpene alcohol sulfates are prepared with chlorosulfonic acid in a basic medium. The sulfates can be further converted to sodium, potassium, or lithium salts. The lithium salts have excellent water solubility and are considered superior to salts with other metals.
[0006] 2. Triterpene phosphates. Phosphate ester derivatives are very attractive candidates as charge modifiers. The esters can be synthesized using phosphoryl chloride or, alternatively, one of the common phosphorylating reagents used in nucleic acid chemistry. Triterpene alcohol phosphates can be further salted with metal ions or organic bases.
[0007] 3. Triterpene-Amino Acid Conjugates. To expand the diversity of polar modifications, we considered the synthesis of conjugates with amino acids that can be acidic, basic, or polar in nature. Lysine and arginine are particularly attractive for this purpose. Moreover, since amino acids form convenient handles for further modifications, this idea can be extended to peptides with affinity for specific receptors.
[0008] 4. Triterpenes conjugated with PEG polymers. PEG technology has been successfully applied in drug formulations to improve drug bioavailability. PEG polymers have the property of forming a polar surface that wraps around drug molecules. PEG carboxylic acids with molecular weights between 500 and 4,000 daltons are sufficient to achieve this goal. It is also rare to employ PEG polymers larger than 4,000 daltons. PEG conjugates are known to degrade gradually in vivo, releasing the core compound.
[0009] 6. Triterpenes conjugated with carbohydrates. Carbohydrates can increase the solubility of selected triterpenes and can be used as guide molecules for intracellular transport mechanisms. The usefulness of this modification may depend more on the latter than on the solubility in aqueous media.
[0010] 7. Encapsulation of triterpenes in complexes with cyclodextrins. This approach is based on modifiers that do not bind covalently to the triterpene. Known examples include ginsenosides, the biologically active triterpenes from ginseng, which have been successfully incorporated into β-cyclodextrin.
[0011] 8. Derivatization of selected triterpenes with dicarboxylic acids. Glutaric and succinic acids are readily available. Conversion of the alcohol group of a triterpene to a carboxylic acid may improve solubility or serve as a convenient handle for the introduction of more polar moieties.
[0012] In one embodiment, activity was demonstrated with euphorin succinate (MDA-MB-231). Inhibitory activity began at doses below 30 ug / ml, with an IC50 of around 17 ug / ml. This is a triple-negative breast cancer cell line.
[0013] According to the present invention, inhibition of cytokines is confirmed and supports the treatment of COVID-19.
[0014] Working Example Cell viability assay Assessment of viability by MTT assay of MDA-MB-231 cells after incubation with Eu-succ. Cells were incubated with test substances (Eu-succ: 0.01-100 μg / mL) for 24 h, and then cell viability assay was performed by MTT method. The vertical axis shows the mean ± standard error of three different tests. The test substance showed IC50: 17.22 μg / mL (10.53-27.28 μg / mL). Viability was calculated relative to the vehicle group (RPMI1640 medium supplemented with 1% DMSO).
[0015] Feasibility study - MTT Assessment of viability by MTT assay of A549, Caco-2, MDA-MB-231 and THP-1 cells after incubation with test substances ST-160.1 and ST-160.2. Cells were incubated with test substances (ST-160.1 and ST-160.2; 1-30 μg / mL) for 24 h, after which cell viability assay was performed by MTT method. Assays were performed in duplicate. (A) ST-160.1 and (B) ST-160.2. Viability was calculated relative to vehicle group (DMEM or RPMI1640 medium supplemented with 1% DMSO).
[0016] Assessment of viability by MTT assay of A549, Caco-2, MDA-MB-231 and THP-1 cells after incubation with test substances ST-160.3 and ST-160.4. Cells were incubated with test substances (ST-160.3 and ST-160.4; 1–30 μg / mL) for 24 h, and then cell viability assays were performed by the MTT method. Assays were performed in duplicate. (A) ST-160.3 and (B) ST-160.4. Viability was calculated relative to the vehicle group (DMEM or RPMI1640 medium supplemented with 1% DMSO).
Claims
1. Use of tetracyclic triterpene 3-ol derivatives with protein kinase activity, including PKC isoforms, in the treatment of cancer, inflammation, and pain.
2. Use of a substance according to claim 1 having the general formula as shown in FIG. 1, wherein R may be a sulfate, a phosphate or a phosphate ester, an amino acid or a peptide, a PEG polymer or other hydrophilic polymer, a dicarboxylic acid or an ester thereof, and R1-8 belong to moieties such as hydrogen, methyl, hydroxyl, ether, hydroxymethyl and its esters or ethers, and a carboxyl group and its esters.
3. 2. Use of the substance according to claim 1 in the form of a pharma- ceutically acceptable salt or derivative, formulation, or cyclodextrin complex, or encapsulated in a bioerodible polymer matrix.