Background and overview[1][2]
Ubenimex is a low-molecular dipeptide compound isolated from the culture medium of Streptomyces oleifera in 1976 by Japanese scholar Hamao Umezawa. Its chemical name is N-[(2S, 3R) -3-Amino-2-hydroxy-4-phenylbutyryl]-L-leucine is in the form of white crystalline powder, easily soluble in glacial acetic acid, and slightly soluble in water or methanol. It was officially launched in Japan in 1987 as an anti-cancer auxiliary drug with immunomodulatory function. The domestic trade name is Baishixin. It is a national second-class new drug and was launched in 1998. It was developed by Sichuan Antibiotic Industry Research Institute and Zhejiang Puluokangyu Pharmaceutical. Ltd., and the tablets and capsules currently on the market are bioequivalent. Adverse reactions that occurred in the clinical trials of Ubimax mainly included rash, erythema, and gastrointestinal reactions. Most of the symptoms will disappear on their own after continuing to take the drug or stopping the drug. As an aminopeptidase inhibitor, ubenimex has a certain anti-tumor effect in inhibiting the growth of tumor cells, inhibiting tumor invasion and metastasis, and exerts a significant synergistic anti-cancer function when used in combination with other drugs. It plays a regulatory role that cannot be ignored in terms of immunity.
Target[1]
Existing research shows that the main inhibitory target of ubenmax is aminopeptidase. This enzyme degrades the extracellular matrix and basement membrane by hydrolyzing the amino acids at the amino terminus of the peptide, thereby promoting tumor metastasis. Expression is upregulated in aggressive, rapidly proliferating malignant tumors. Ubimax can inhibit the activity of aminopeptidase by binding to the Zn2+ and hydrophobic active center of aminopeptidase. At present, among the dozen or so aminopeptidases inhibited by ubenimex, the most studied ones are aminopeptidase N (aminopeptidase N, also known as CD13) and aminopeptidase B, among which CD13 is the most important. In addition to the joint hydrolysis of the above-mentioned aminopeptidases affecting tumor invasion and metastasis, CD13 is also related to blood vessel formation, viral receptors, antigen presentation, and regulatory cytokines. The highly efficient probe with ubenimex activity developed in 2008 can more broadly detect the functional properties of metalloaminopeptidase in different biological systems.
In addition to targeting aminopeptidase, another inhibitory target of ubenmax is leukotriene A4 (LTA4) hydrolase. Glu-296 is an important structural determinant in the binding process of the two. In addition, some researchers have found that ubenimex is also a competitive inhibitor of tumor dipeptidase. This enzyme is isolated and purified from mouse ascites tumor cells. Different from the binding method of aminopeptidase, ubenimex binds to The dipeptidase binding process is slow but not tight. In summary, the main targets of ubenimex are aminopeptidase and LTA4 hydrolase.
Mechanism of action[1]
In the early days, it was believed that the anti-tumor activity of ubenmax in the body is to kill tumor cells by affecting the body's immune response, such as activating immune cells macrophages, degrading and inactivating immune active factors, activating granulocytes and NK cells, Promote anti-tumor non-specific immune responses, etc. Subsequently, a large number of studies found that even without immune response, ubenimex still has anti-tumor effects. Therefore, the anti-tumor effects of ubenimex include direct and indirect anti-tumor effects through immune regulation. effect.
1. Cell proliferation inhibition and apoptosis induction
In 2000, it was discovered that the proliferation of human promyelocytic acute leukemia cell line HL60 cells could be inhibited by ubenimex. Ubimax can also directly induce apoptosis in chronic myelogenous leukemia K562 cells and inhibit cell proliferation. This process depends on the activation of caspase-3 and is also related to the phosphorylation of mitogen-activated protein kinase (MAPK) and glycogen synthase kinase-3β (GSK-3β) and the inhibition of cyclin D1 expression. In normal colorectal cancer cells, the promoter of the death-associated protein kinase (DAPK) gene is in a methylated state. However, after the cells were treated with ubenimex, the promoter of the gene was reversed to a demethylated state, and both DAPK mRNA and protein were in a methylated state. Expression is restored, cell proliferation is inhibited and apoptosis occurs. Other literature shows that ubenimex and its analogues bind to the BIR3 domain, causing apoptosis inhibitory factor 1 to be automatically ubiquitinated and degraded, ultimately leading to apoptosis of tumor cells. In addition, it was found that in human solid tumor cells, ubenimex can promote Fas agonistic antibody CH11 to induce apoptosis of tumor cells (human cervical cancer cells HeLaS3 and fibrosarcoma cells HT-1080) without causing tumor cell apoptosis, and enhance Activation of caspase-8. Currently, among the ubenimex analogs that have been successfully synthesized, LYP is more effective in inhibiting the growth of ovarian cancer ES-2 transplanted tumors in vivo. LYP2 can inhibit the growth of six types of tumor cells, including HL60, A549 and PLC, with IC50 The values are lower than those of ubenimex, and the growth inhibitory effect of LYP3 in ES-2 and HL60 tumor cells is also better than that of ubenimex. Moreover, the above analogs have better water solubility and biological stability, and may become effective substitutes for ubenimex in future research.
2. Inhibition of invasion and metastasis
A large number of studies have found that in metastatic tumors, ubenimex can inhibit the invasion and metastasis of tumor cells by inhibiting CD13 activity, and it is ubenimex concentration-dependent, and is accompanied by the extension of the survival time of mice. Early scholars found that ubenimex can significantly reduce the expression level of 68kDIV type collagenase protein (active form). The inhibitory effect of ubenimex on tumor cell invasion may be related to its influence on the conversion process of type IV collagenase from latent form to active form. and tumor cells��Intestinal reaction; some individuals have liver damage, and there may be a transient slight increase in serum aspartate aminotransferase (AST), which disappears during the medication or after stopping the medication; occasionally headaches and numbness; visible skin Redness, mild hair loss, rash, itching. Occasionally there is facial swelling.
Notes[2]
The safety for pregnant and lactating women and children has not been established and should be used with caution. Daily doses exceeding 200mg can reduce the number of T cells.
Preparations and Specifications[2]
Tablet: 10mg; capsule: 10mg, 30mg.
Combined medication[1]
Due to the complexity of tumors, traditional treatment methods such as radiotherapy and chemotherapy are not effective in curing some malignant tumors that are difficult to cure and prone to recurrence. Combination drug strategies aimed at improving efficacy will be the general direction for the treatment of cancer in the future. . Ubimax is now used in combination with a variety of chemical drugs to treat tumors, with obvious effects. Recent research
It shows that when ubenimex is combined with the chemotherapy drugs 5-fluorouracil, doxorubicin, leucovorin, etc. to treat late-stage recurrent liver cancer caused by hepatic artery embolism (especially small liver cancer with tumor diameter <3cm and 3~5cm), It will enhance cell apoptosis and intracellular reactive oxygen species levels, and the therapeutic effect is better than chemotherapy alone. For advanced gastric cancer, when combined with the chemotherapy regimen of Tegio and oxaliplatin, ubenimex can reduce the patient's serum MMP2 and MMP9 levels and enhance the anti-tumor effect. Ubumax can also enhance the inhibitory effect of all-trans retinoic acid on the proliferation of acute promyelocytic leukemia cells and its ability to induce cell differentiation. In addition, ubenimex can significantly improve the inhibitory effect of cisplatin on the growth of A549 cells, and may become a sensitizer for cisplatin treatment of CD13+ non-small cell lung cancer.
Preparation[3]
The preparation method of ubenmax is as follows:
1) Add L-leucine benzyl ester p-toluenesulfonate, HOBt, and 10 times (V/m) ethyl acetate into the reaction kettle in sequence. After the feeding is completed, stir and cool down continuously, and slowly add DCC ethyl acetate. The ester solution is added to the process system at a temperature between 0-5°C. After the feeding is completed, the system reacts at 15-20°C for 4 hours. TLC tracks the reaction endpoint (ethyl acetate: petroleum ether = 2:1, UV 254nm) to obtain an activated ester solution of L-leucine benzyl ester p-toluenesulfonate. ;
2) Add (2S, 3R)-3-benzyloxyformamido-2-hydroxy-4-phenylbutyric acid and 10 times (V/m) 10% sodium bicarbonate solution into the reaction kettle in sequence , stir until completely dissolved, cool down to 10-15°C, add the activated ester solution of L-leucine benzyl ester p-toluenesulfonate dropwise with rapid stirring, and react at 15-20°C for about 4 hours after the dropwise addition is completed. TLC tracking reaction end point (ethyl acetate:petroleum ether=2:1, UV 254nm);
3) Filter and separate the liquids. Extract the water phase with 7 times and 2 times (V/m) of ethyl acetate in sequence. Combine the ethyl acetate phases. Wash the ethyl acetate phase with 6 times of 0.5wt% hydrochloric acid. Wash once with 6 times 5wt% saturated sodium bicarbonate and once with 6 times purified water; dry the ethyl acetate solution over anhydrous sodium sulfate for 2 hours, filter and concentrate to dryness, add 5 times (V/m) petroleum to the concentrate Ether, stir for 2 hours to disperse, filter, and dry at 40-45°C to obtain N-[(2S, 3R)-3-benzyloxyformamido-2-hydroxy-4-phenylbutyryl]-L-leucine, Yield 94-98%;
4) Combine N-[(2S, 3R)-3-benzyloxycarboxamido-2-hydroxy-4-phenylbutyryl]-L-leucine, Pd/C (30%) at room temperature ) is dissolved in 15 times (v/m) glacial acetic acid, stir and mix evenly and then add it to the reactor. Replace nitrogen and hydrogen three times in sequence. Stir and react at 1.5-2MPa until no hydrogen is absorbed;
5) The filtered filtrate is concentrated to dryness at 80°C and -0.1MPa. Add 10 times (v/m) acetone to the concentrate, stir and disperse for 4 hours, filter, and blow dry at 40-45°C to obtain black Benmex acetate;
6) Dissolve ubenimex acetate in hydrochloric acid solution, cool the solution to between 0-5°C, and adjust the pH value of the solution to 5-6 with 15wt% ammonia water. Crystallize at 0-5°C 5h, filter, and the solid is pulped with acetone for 3h, filtered, and dried at 40°C to obtain the finished product of ubenimex, with a yield of 90.5%.
Main reference materials
[1] Research progress on the anti-tumor effect of ubenimex
[2] Practical Drug Handbook
[3] CN201710552900.0 Preparation method of ubenimex
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