The influences of alkaloids of Chelidonium majus L., Colchicum autumnale L, Catharanthus roseus (L.) G.Don and Vinca minor L. on malignant neoplasms, the review of modern researches

: natural alkaloids of Chelidonium majus L., Colchicum autumnale L. and Catharanthus roseus (L.) G.Don are anticancer agents. Some of them, such as colchicine, vincristine and vinblas - tine, are used in modern medicine, as chemotherapy medicines against malignant neoplasms, some of them are effective supplement to conventional methods or works to prevent cancer onset (chelidonine, sanguinarine, chelerythrine, protopine and allocryptopine). The effect of mitotic poisons that are al - kaloids of Colchicum autumnale colchicine, Catharanthus roseus vincristine and vinblastine against malignant neoplasms began to be studied in the last century, the fact of mitotic spindle violation is given in a large number of works. However, the mechanisms of apoptosis

Among the medical plants containing alkaloids, the most noteworthy are Chelidonium majus L, Colchicum autumnale L, Catharanthus roseus (L.) G.Don and Vinca minor L. The effect of mitotic poisons that are alkaloids by Colchicum autumnale colchicine and Catharanthus roseus vincristine and vinblastine against malignant neoplasms began to be studied in the last century, the fact of mitotic spindle violation is given in a large number of works.However, the mechanisms of apoptosis under their influence have been little investigated.The alkaloids of Chelidonium majus and Vinca minor have been much less studied, both in clinical studies and experimental ones, including insufficiently researched their antiproliferative action, the ability to cause apoptosis and its possible mechanism.
The research of apoptosis mechanisms caused by natural antitumor agents, will allow creating more effective and saving medicines based on the active ingredients of plant raw materials in the future.

Aim
To analyze the influence of the chelidonine, sanguinarine, chelerythrine, protopine and allo-cryptopine contained in Chelidonium majus; colchicine contained in Colchicum autumnale; vincristine and vinblastine contained in Catharanthus roseus; vincamine contained in Vinca minor on malignant neoplasms and the mechanisms of these agents` effects reflected in modern scientific literature.

Materials and methods
Analytical review of Ukrainian and foreign literature for the period 2002-2023, which is dedicated to the effect of Chelidonium majus, Colchicum autumnale, Catharanthus roseus, Vinca minor and/or their main alkaloids on malignant neoplasms and the mechanisms of such an influence with the help of medical database PubMed.
According to Habermehl, Kammerer & Handrick, (2006) drug Ukrain, which included alkaloids of Chelidonium majus chelidonine, sanguinarine, chelerithrine, protopine and allocryptopine, is an inducer of malignant cells apop tosis.The drug caused depolarization of the mitochondrial membrane and caspases activation.This process is shown in Figure 1.Caspase-8 and FADD (Fas-associated Death Domain) are two important signaling molecules of the death receptors apoptosis pathway.It was reported that Ukrain-induced apoptosis did not require the presence of caspase-8 and FADD, expression of the caspase-8 inhibitor cFLIP-L (cFLIP -cellular FADD-like interleukin-1 converting enzyme (FLICE) inhibitory protein) or resistance to the death receptor ligands failed to inhibit Ukrain-induced apoptosis.This indicates a signaling pathway independent of the death receptor (Habermehl et al, 2006).At the same time, broad-spectrum caspase inhibitor zVAD-fmk blocked drug-induced cell death.The authors also noted the participation of Bcl-2 (B-cell lymphoma/ leukemia-2), which are regulatory proteins exactly of mitochondrial pathway of apoptosis (Habermehl et al, 2006;Papaliagkas, Anogianaki & Anogianakis, 2007).Both pathways, extrinsic (death receptors) and intrinsic (mitochondrial) with the expression of caspase-8 inhibitor cFLIP and the resistance to death receptors are shown in the Figure 2.  The drug Ukrain, produced from Celandine, in combination with conventional antitumor treatment increased the survival of patients with pancreatic cancer (Gansauge et al, 2002).Its therapeutic effect on a number of malignant neoplasms is noted, while the methodological basis of most clinical studies was insufficient (Ernst & Schmidt 2005).
Among the alkaloids of Chelidonium majus, chelidonine is most studied.It induces apoptosis of cancer cells (Havelek, Seifrtova & Kralovec 2016;Noureini & Esmaili, 2014;Noureini & Wink, 2009;Paul, Bishayee & Ghosh, 2012;Камінський В.О., 2006;), including human lymphoma cells and works against this neoplasm together with other alkaloids, which are sanguinarine, chelerithrine and coptisine (Камінський В.О., 2006).Chelidonine also induces apoptosis of MCF-7 breast cancer cell line (MCF-Michigan Cancer Foundation, MCF-7 is breast adenocarcinoma cell line) (Noureini & Esmaili, 2014), HeLa cells (HeLa is cervical cancer cell line), (Paul et al, 2012), leukemic Т-cells (Havelek et al, 2016), human ovarian cancer cells (Shen, Lee & Joo, 2022) of chelidonine cytotoxic action, it is noted that chelidonine induces apoptosis at low doses in MCF-7 breast adenocarcinoma cells, but at high doses, it causes autophagy of this type of cancer cells (Noureini & Esmaili, 2014).There are studies of apoptosis`s mechanisms caused by chelidonine alone.Камінській (2006) notes that the induction of apoptosis was carried out at mitochondrial level.The author also suggests that the induction of lymphoma cells apoptosis under the influence of chelidonine can be carried out with the involvement of caspase-9 and caspase-8, the last one takes part in receptor-mediated pathway of apoptosis (Камінський В.О., 2006).Caspases participate in two different pathways of apoptosis: extrinsic, mediated through death receptors and subsequent activation of caspase-8, and intrinsic, mitochondrial pathway through activation of caspase-9 (Wen, Lin & Liu 2012).At the same time, caspase-8, which is formed at extrinsic pathway, can also activate the intrinsic pathway through Bid (Bid is proapoptic BH3 domain bсl2), that is, through a proapoptic member of the protein family bcl-2.Moreover, there are ways for caspase-8 activation without death receptors (Wen et al, 2012).These apoptic ways are shown in Figure 3. Taking into consideration the most scientific data, we suggest these mechanisms are to be the most probable.The participation of both caspase-8 and caspase-9 is confirmed by Havelek et al (2016).Data by Камінський (2006) and Havelek et al (2016) partially contradict the data by Habermehl et al. (2006), who proved the insignificance of caspase-8 expression in apoptosis under the influence of drug Ukrain.
In favor of mitochondrial pathway of the effect of chelidonine, data on the disappearance of the mitochondrial membrane potential indicate (Havelek et al, 2016;Paul et al, 2012).Cell cycle arrest (Havelek et al, 2016;Noureini & Wink, 2009;Paul et al, 2012), DNA fragmentation (Paul et al, 2012) under the influence of chelidonine and inhibition of telomerase in submicromolar concentrations of chelidonine (Noureini & Esmaili, 2014) are also noted.These data support Noureini & Wink, (2009), who reported that low doses of chelidonine caused a decrease in telomerase activity.
The study of signaling pathways in HeLa cells showed that chelidonine could effectively induce apoptosis due to the expression of genes encoding p38, p53 and other proapoptic genes and suppressing the expression of Protein kinase B (AKT), phosphatidylinositol 3-kinases (PI3K), Janus kinase 3 (JAK3), signal transducer and activator of transcription 3 (STAT3), oncoproteins E6 and E7, and anti-apoptotic genes (Paul et al,

2012)�
Autumn crocus (Colchicum autumnale L), Colchicaceae family.All parts of the plant contain alkaloids: colchicine and colchamine.Colchicine has significant anti-proliferative activity (Li, Hu & Pu, 2022), it inhibited sufficiently proliferation and induced apoptosis by modulating the expression level of several genes and carried out anticancer effect on human breast adenocarcinoma MCF-7 and mouse breast cancer cell line 4T1 (Adham Foumani, Irani & Shokoohinia, 2022).The authors suggest that colchicine may be a potential candidate for the prevention and treatment of breast cancer.A serious drawback of colchicine, which limits its use in effective doses, is toxicity (Lin, Yeh & Huang, 2021), however, a currently synthesized colchicine-magnolol hybrid inhibited the growth of lung carcinoma cells (Li et al, 2022).Colchicine works primarily by inhibiting microtubule polymerization (Dhyani et al, 2022) which in turn, affects numerous cellular processes, namely: maintenance of shape, signaling, division, migration, and cellular transport (Angelidis, Kotsialou & Kossyvakis, 2018).In addition to its effect on microtubule polymerization, which is anti-proliferative, colchicine also induces apoptosis.Chen et al, (2012) believe that apoptosis under the action of colchicine is carried out through the intrinsic, i.e. mitochondrial, pathway.A decrease of the mitochondrial membrane potential, activation of caspase-3 and -9, an increase of Bax (BCL2 associated X protein, apoptosis regulator) and a decrease of Bcl-2 were shown (Chen et al, 2012).According to these data, this mechanism is similar to the one shown in Figure 3 for celandine alkaloids.
The mechanism of pink periwinkle alkaloids is the following: inhibition of tubulin polymerization in low doses, tubulin polymerization is necessary for the formation of the mitotic spindle during the metaphase of the cell cycle; in high doses, pink periwinkle alkaloids cause cell cycle arrest and apoptosis.(Arora & Menezes, 2021).The mechanism of action on microtubules is similar to that of colchicine, with tubulin having three binding domains: paclitaxel, Catharanthus roseus alkaloids and colchicine domains.(Cheng & Feng (2020) Small periwinkle (Vinca minor L.), Apocynaceae Family Vincamine, an alkaloid of Vinca minor, is used as a dietary supplement and vasodilator.Al-Rashed, Baker & Ahmadl, (2021) have demonstrated for the first time the possibility of using vincamine as an antitumor agent on the adenocarcinoma cell line A549 of human alveolar basal cell epithelium.Vincamine stimulated caspase-3-dependened apoptosis and reduced mitochondrial membrane potential, stimulated the release of cytochrome C. The non-toxicity of vincamine was shown on non-tumorigenic cell lines -BEAS-2B (human bronchial epithelial cell line) and 3T3-L1 (cloned cell line from a mouse embryo) (Al-Rashed et al, 2021).The release of cytochrome C and decreased mitochondrial membrane potential, which were researched by Al-Rashed et al. (2021), are indicators of the intrinsic signaling pathway of apoptosis.This way is shown in Figure 1.It is significant that one study proved the apoptosis of malignant cells and the absence of toxicity of the drug for healthy cells.

Conclusions
According to the analysis of the literature of recent years, the cytotoxic and anti-proliferative effects of natural alkaloids of Chelidonium majus chelidonine, Colchicum autumnale colchicine, Catharanthus roseus vincristine and vinblastine and Vinca minor vincamine on malignant cells can be considered proven.
Colchicine caused apoptosis in high doses, chelidonine, on the contrary, caused apoptosis of malignant cells in relatively low doses; in large doses it caused autophagy.
The signaling pathways of apoptosis mechanisms of malignant cells under the influence of chelidonine, colchicine, vincristine, vinblastine and vincamine are much less studied and require additional research.Most of the results support the mitochondrial pathway, but there is a view in favor of the receptor-mediated pathway.

Figure 1 .
Figure 1.The probable scheme of malignant cells apoptosis under celandine alkaloids with the participation of intrinsic (mitochondrial) pathway.Depolarization of mitochondrial membrane causes cytochrome C releasing from mitochondria, which, together with Apaf-1 (Apoptotic peptidase activating factor 1) and procaspase -9 form apoptosome. Apoptosome activates caspase-9.the last can also be activated by procaspase-9.Activated caspase-9 activates caspase-3, which destroys inhibitor of DFF (DNA fragmentation factor).DFF activates CAD (caspase-activated DNase).CAD is an enzyme destroying DNA, which in turn leads to the final stage of apoptosis.

Figure 2 .
Figure 2. The probable scheme of malignant cells apoptosis under celandine alkaloids with the participation of extrinsic pathway and intrinsic (mitochondrial) pathways with the expression of procaspase-8 inhibitor and death receptors resistance.The expression of procaspase-8 inhibitor (FLIP, green color) causes the impossibility of caspase cascade activation from procaspase-8 to caspase-7.The same result takes place under death receptor resistance.The failed ways are shown with red.In spite of it, the apoptosis is possible.Apaf-1 -Apoptotic peptidase activating factor 1 DFF -DNA fragmentation factor) CAD -caspase-activated DNase)

Figure 3 .
Figure 3.The probable scheme of malignant cells apoptosis under celandine alkaloids with the participation of extrinsic pathway and intrinsic (mitochondrial) pathways without death receptors participation.Apaf-1 -Apoptotic peptidase activating factor 1 DFF -DNA fragmentation factor) CAD -caspase-activated DNase) Bid, Bax, Badproapoptic proteins

com Ukrainian Scientific Medical Youth Journal
. Apoptosis is not the only way Ukrainian scientific medical youth journal, 2023, Issue 4 (142) http://mmj.nmuofficial.com