Nandkumar Shinde College of Pharmacy, Aghur, Vaijapur 423701 Dist.-Aurangabad, Maharashtra
Neem, or Azadirachta indica, is a plant that is native to the Indian subcontinent and is a member of the Meliaceae family. Because of its many biological benefits, the Neem tree is also known as the village pharmacy. There are several applications for every part of the Neem tree, including its bark, leaves, sap, fruit, seeds, and twigs. They are commonly used to treat a variety of infections and skin conditions Azadirachtin and nimbolide are among the few bioactive components in neem that have been studied extensively, but research on a great number of additional bioactive components is warranted. The key anticancer effects of neem components on malignant cells include inhibition of cell proliferation, induction of cell death . Neem’s anticancer qualities, which have been previously investigated, include its capacity to alter the tumor environment, boost host monocyte cytotoxicity, and inhibit tumor cell growth. The goal of the current review was to examine and compile information regarding Azadirachta indica’s potential benefits for treating cancer.
The plant product or natural products play a significant role in the prevention and treatment of diseases by enhancing antioxidant activity, inhibiting pathogen growth, and modulating genetic pathways. The therapeutics role of number of plants in diseases management is still being enthusiastically researched due to their less side effect and affordable properties. It has been accepted that drugs based on allopathy are expensive and also exhibit toxic effect on normal tissues and on various biological activities. It is a largely accepted fact that numerous pharmacologically active drugs are derived from natural resources including medicinal plant.[1]Many infectious, metabolic, and cancerous disorders are treated with neem (AzadirachtaIndica) components in Ayurveda,Unani, homeopathy, and contemporary medicine.[2] Neem, or Azadirachta indica, is a plant of the Meliaceae family that has long been known for its therapeutic benefits. It grows in tropical and semi-tropical areas of the world, and its various parts—including seeds, leaves, flowers, and bark—are utilized extensively for a variety of uses . It also elaborates a wide range of chemically diverse and structurally complicated biologically active molecules. [3]Proteins and carbohydrates are the two main families of isoprenoids and non-isoprenoids, which comprise the more than 150 chemicals that have been isolated from various neem components. Additionally, it contains sulfurous chemicals, polyphenolic components such flavonoids and their glycosides Neem contain flavonoids. Increase the flavonoids consumption then decrease the cancer risks. dihydrochalcone, coumarin and tannins, and aliphatic compounds, and isoprenoid compounds, such as diterpenoids and triterpenoids that contain protomeliacins, limonoids, azadirone and its derivatives, gedunin and its derivatives, vilasinin type of compounds, and c-secomeliacins like nimbin, salanin, and Nimbidin . Its Taxonomic position is as follows[4]
Order Rutales Suborder Rutineae.
Family Meliaceae (mahogany family). Subfamily Melioideae.
Tribe Melieae Genus Azadirachta.
Species Azadirachta indica.
Neem leaf ethanolic extracts and neem liminoids are the most widely used preparations or extracts made from the seeds, bark, fruits, leaves, and other parts of the neem tree. These neem components’ biological qualities have been evaluated in both non-neoplastic and other neoplastic circumstances. The effects of neem’s various components are not well understood. [5,6]
1. LIMONOIDS:
At least nine neem limonoids have so far shown the capacity to inhibit the growth of insects, affecting a variety of species that include some of the most dangerous agricultural and human health pests. Although new limonoids are still being found in Neem, the most well-known and, at least for the time being, most important ones include azadirachtin, salannin, meliantriol, and nimbin.
2. MELIANTRIOL:
another feeding inhibitor, can stop insects from eating at very low doses. Neem’s traditional use for controlling insects on Indian crops was first scientifically proven when it was shown to be effective in preventing locusts from nibbling on crops.
3. SALANNIN :
Salannin is a third triterpenoid that has been identified from Neem. Research shows that while this chemical has no effect on insect molts, it also effectively prevents feeding. In both lab and field studies, the migrating locust, California red scale, striped cucumber beetle, houseflies, and Japanese beetle have been significantly repelled.
4. NIMBIDIN AND NIMBIN :
It has been discovered that two more Neem constituents, nimbin and nimbidin, exhibit antiviral properties. They impact the vaccinia virus, the fowl pox virus, and the potato virus X. Perhaps they will make it easier to control these and other viral illnesses that affect cattle and crops. When neem seeds are extracted using alcohol, the main ingredient in the bitter principles that are produced is nimbidin. It is present in significant amounts, roughly 2% of the kernel
• THE ACTIVE COMPOUNDS OF AZADIRACHTA INDICA :
Sometimes known as neem, have a therapeutic effect in managing health because of their abundance of different kinds of substances. The most significant active ingredient is azadirachtin, followed by sodium nimbinate, gedunin, salannin, quercetin, nimbolinin, nimbin, nimbidin, and nimbidol. Nimbin, nimbanene, 6-desacetylnimbinene, nimbandiol, nimbolide, ascorbic acid, n- hexacosanol, amino acid, 7-desacetyl-7-benzoylazadiradione, 7-desacetyl-7-benzoylgedunin, 17- hydroxyazadiradione, and nimbiol are among the compounds found in leaves [7,8]. Neem seeds
• MEDICINAL PROPERTIES OF NEEM :
For over 2000 years name has been used in traditional medicine in India to trade range of ailments
Long disease and condition affected by these smallpox diabetes fever tumours malaria and ulcer Various properties including
Anti-inflammatory Anti ulcer
Antimalarial antibacterial
antioxidant together with antiseptic Astringent
emollinment
Anti helminthic’s an
insecticides hence neem shows Various pharmacological properties Antioxidant antimicrobial an anti cancer potential of Neem This include
1. Ripe an unripe fruits
2. Cool and fresh fruits
3. Fruit epicarp 4.Leaves
5. Flowers and steam bark and roots
• EXTRACTION PROCESS :
Despite being present throughout the tree, the most concentrated and easily accessible bioactive chemicals are located in the seed kernels. They are made by extracting different parts of the kernels and, to a lesser degree, the press cake. In organic solvents like hydrocarbons, alcohols, ketones, or ethers, the active components are freely soluble, despite being very weakly soluble in water.[9]
EXTRACTION OF WATER :
The most straightforward method (and the one used the most nowadays) is to grind or crush the kernels and then use water to extract them. For instance, a cloth bag hanging in a barrel of water could be used to steep them for the entire night. This method is less efficient than simply pouring the water into the bag and gathering the extract as it comes out, for reasons that are still unknown. Without undergoing any additional changes, the final crude suspension is suitable for field use. As a sprayable emulsion, it can also be filtered. In Third World villages, this is the most promising strategy. Twenty to thirty kilograms of neem seed may typically treat one hectare when water extraction is used, according to estimates. Currently, the yearly seed crop from However, because the active components are so poorly soluble in water, a large amount of water must be used. Typically, the quantities used are approximately 500 g of kernel steeped in 10 liters of water. Water extracts of powdered neem leaves are also extremely beneficial. Because Neem is an evergreen, it is available throughout the year.
EXTRACTION OF HEXANE :
If the kernels are grated and soaked in the solvent hexane, just the oil is extracted. The oil isn't regarded as an active pesticide. However, current discoveries suggest that it is an extremely intriguing material, which in some situations can be used to kill the eggs of many types of insects, the larvae of mosquitoes, and other stages of certain pests (such as leafhoppers).The residue left after the hexane extraction still retains the major active limonoid components, and additional water or alcohol extractions generate huge amounts of them that are pure and free of oil.
EXTRACTION OF ALCOHOL :
The simplest method for creating concentrated neem-based pesticidal compounds is alcohol extraction. In alcohol-based solvents, limonoids are very soluble. Ethanol, but occasionally methanol, is used to soak the grated kernels. Active component yields range from 0.2 to 6.2 percent.5. Neem chemicals are not very soluble in water, despite the fact that water extracts work well as pesticides; the alcohol extracts are roughly 50 times more concentrated. They might have up to 100,000 parts per million (ppm) of azadirachtin.
• FORMULATION :
As mentioned, a crude extract is the most basic form of neem insecticide. However, a number of changes can be made for more advanced use. Neem extracts can be transformed into granules, dust, wettable powders, or emulsifyable concentrates using these sophisticated formulas. For easier application against skin conditions, aqueous extracts can also be made with soap. In other formulations, the neem components themselves may be chemically modified or chemicals may be added. These adjustments could be made to improve repeatability and shelf stability, as well as to make handling or process scaling easier. Additionally, they might lessen phytotoxicity, which harms delicate plants. Additives that prevent UV deterioration are one particularly useful kind. These consist of para-aminobenzoic acid (PABA), lecithin, and sesame oil.
• ADDITIVES:
Neem extracts’ potency can be increased ten to twenty times by combining them with other substances. These so-called “promoters” include piperonyl butoxide, sesame oil, and pyrethrins, a pesticide primarily derived from chrysanthemum flowers . They are employed to achieve a faster kill. Synthetic pesticide combinations can also be effective since they provide a quick “knockdown” to neem’s capacity to inhibit the insect population’s subsequent return. The insect-killing bacterium Bacillus thuringensis (Bt) can even increase the efficacy of neem extracts, creating a multipurpose insecticide.
• CHEMOTHERAPEUTIC EFFECT OF NEEM :
The effects of neem on chemotherapy Neem bark and leaf aqueous and alcohol extracts are powerful antimalarials in West Africa, India, Burma, and other places, especially against types of malaria that are resistant to chloroquine. Gedubin, one of the active ingredients, significantly reduced malaria and was just as effective as quinine. Red blood cells’ (RBCs’) redox state on parasites may be the method. Neem extracts reduced the oxidized cells to eliminate the malaria parasite, which is produced by the plasmodial parasite. [4] Additionally, in Germany and India, neem leaves and barks are used as an active ingredient in dental paste due to its potent antibacterial qualities. Aqueous leaf extract, on the other hand, has laxative properties through increased bowel movements.
Components of NEEM diminish cellular oxidative stress, cause apoptosis and other types of cell death, and inhibit growth (Fig. 1). In the carcinogen-induced hamster buccal pouch (HBP) model, neem leaf extract (NLE) changes the expression of genes controlling many cellular functions [9]. Angiogenesis and metastasis are significantly influenced by the tumor microenvironment. Tumor cells have the capacity to alter their immediate surroundings, often known as the microenvironment, which promotes angiogenesis, cell invasion, and inflammation [10,11,12]. As a result, the tumor microenvironment is crucial to the development and spread of malignancies. Curiously, neem components seem to alter the tumor microenvironment through a variety of ways, such as increased immune system cytotoxicity and reduction of angiogenesis. For instance, an in vitro study indicates that human migration and proliferationEther, petrol ether, ethyl acetate, and diluted alcohol are among the solvents used to make neem extracts. Consequently, the range of bioactive components and the proportion of each component in the extract differ according to the extraction method. This variation is probably going to have an impact on the precision of data interpretation and comparisons between other neem extract investigations. Since the majority of research investigates the effects of neem on cancer using solvent-extracted mixtures of its constituent parts, little is known about the roles played by each component. Azadirachtin and nimbolide are two of the few bioactive ingredients that have been thoroughly investigated. A secondary metabolite of neem, azadirachtin, is mostly found in neem seeds [13,14]. Because its structure is so intricate, the initial synthesis
fig:1
• NEEM COMPONENTS INHIBIT CANCER CELL PROLIFERATION :
Components of Neem prevent the growth of cancer cells. One of the primary characteristics of cancer is the unchecked growth and proliferation of cancer cells, which also contributes significantly to the development of tumors and cancer metastases [15,16]. Consequently, a common characteristic of many chemopreventive and therapeutic medicines is their ability to limit the proliferation of tumor cells. Neem extracts inhibit the growth and multiplication of tumor cells by interfering with the progression of the cell cycle. For instance, NLE exhibits proliferation-inhibitory actions in prostate cancer cells [17, 18], while neem seed oil suppresses the growth of HeLa cervical cancer cells [19]. Remarkably, the anti-proliferative actions of NLE in prostate cancer cells are not modulated by androgen dependence. Neem extract, for instance, inhibits the growth of prostate cancer cells that are androgen-dependent as well as those that are not [17,18]. Given that prostate cancer cells are androgen-refractory Patients with recurrent prostate cancer may benefit therapeutically from therapy with active neem components because androgen- refractory prostate cancer cells are more resistant to apoptosis and cause prostate cancer recurrence. Neem’s anti-proliferative actions are consistent in both estrogen-dependent and -independent breast cancer cells, much like its lack of androgen dependency [19].A complex network of regulatory proteins, including cyclins, cyclin-dependent kinases (CDKs), CDK inhibitors (CKIs), cell cycle checkpoint proteins, and transcription factors like E2F, closely regulate the course of the cell cycle [12,20,21]. Numerous target proteins have been found in research on how Neem or its constituents affect the cell cycle and tumor cell proliferation. For instance, azadirachtin treatment of HeLa cells results in the production of CKI p21 and a decrease in cyclin B and cyclin D1 levels, which together cause G0/G1 cell cycle arrest [22]. This active neem component causes both G0/G1 and G2/M arrest, along with changes in cyclins, CDKs, and CKIs, according to an analysis of the cell cycle distribution in colon cancer cells treated with nimbolide Additional G2/M cell cycle nimbolide targets Rad17 and CHK2 are additional nimbolide targets for G2/M cell cycle checkpoint proteins . Nimbolide suppresses the growth of HeLa [22], breast cancer [19], choriocarcinoma , lymphoma [23], leukemia, and melanoma cells [23] by interfering with the course of the cell cycle, for which specific mechanisms are unknown. Analogous inhibitory effects on malignant cell growth and proliferation are demonstrated by other identified neem components. Treatment with NLE, for instance, or gedunin produced from neem reduces the growth of ovarian or pancreatic cancer cells, respectively [24]. A bioinformatics investigation revealed that the subgroups of differentially regulated genes triggered by gedunin encode proteins involved in cell cycle regulation and other cellular functions. Surprisingly, gedunin and cisplatin together reduce treated ovarian cancer cell multiplication by as much as 50%.
The clinical significance of the in vitro results is confirmed by in vivo studies of neem extracts or components, which exhibit strong anticancer effects. In the carcinogen 7,12-dimethylbenz[a]anthracene (DMBA)-induced HBP mouse model, NLE inhibits the carcinogenesis process. This is accompanied by an upregulation of cytokeratin and a decrease in proliferating cell nuclear antigen (PCNA), indicating that neem components suppress proliferation and induce differentiation, respectively .
• EFFECT OF NEEM COMPONENTS ON CANCER CELLS :
Neem components’ impact on the demise of cancer cells It has been demonstrated that neem components effectively induce apoptosis and promote autophagy, which results in the death of cancer cells. Extracts from neem seeds and leaves have been tested for their enhanced apoptotic effects in a number of human malignancies, including breast[19], prostate[25], cervical[26] and leukemia[27]. Neem extracts cause a series of molecular events in the oral cavity that correlate with the findings of increased apoptotic cell death. These events include an increase in the expression of Bim, Bax, Apaf-1, caspase 8, caspase 3, and PARP cleavage, according to studies of DMBA-induced hamster buccal pouch oral carcinogenesis models. It is also observed that Bcl-2 expression is suppressed concurrently.[28,29]
• NEEM ALTERS THE ENZYMES THAT BREAK DOWN XENOBIOTICS :
Enzymes that break down xenobiotics are crucial targets for chemotherapy in anticancer treatment. In their method of action against cancer-causing cells, liminoids like nimbolide and azadirachtin, as well as neem leaf extracts, have two distinct functions. They block phase 1 (carcinogen activation) enzymes and simultaneously increase activation of phase 2 (carcinogen detoxification) enzymes. Because of this dual strategy, neem extracts are beneficial for reducing early-stage carcinogenesis.[30]
• NEEM COMPONENTS’ INFLUENCE ON ENZYMES INVOLVED IN METABOLISM :
Xenobiotics, including carcinogens, are metabolized by drug-metabolizing enzymes, which also maintain cellular reduction/oxidation (redox). Their expression and activity are therefore good targets for therapeutic treatments because of these characteristics [31]. Xenobiotics frequently undergo phase I reactions that result in reactive intermediates, which are then further altered by phase II metabolic enzymes. Neem extracts have the ability to manipulate both phase I and phase II metabolizing enzymes, which can result in balanced redox levels and reduced carcinogen genotoxicity. According to the in vitro Ames test, neem oil reduces the mutagenicity of the carcinogens mitomycin and DMBA [32]. DMBA or N-methyl-N’-nitro-N- nitrosoguanidine (MNNG) can cause bone marrow micronuclei, while NLE inhibits this process NLE reduces the occurrence of cancers in animals exposed to various carcinogens, which is consistent with its effect on lowering genotoxicity [33]. A key signaling molecule in the regulation of cell survival and proliferation are cellular reactive oxygen species (ROS), which are primarily generated by mitochondrial respiratory chain reactions [34,35]. However, biological macromolecules like DNA, lipids, and proteins are harmed by an overabundance of ROS. Deregulated metabolism and mitochondrial malfunction can result in increased ROS generation above the normal level, which is linked to aging and chronic illnesses like cancer . Therefore, oxidative stress-induced damage is successfully prevented and the incidence of cancer is decreased by controlling cellular ROS levels and redox equilibrium [36]. Rats fed neem flowers as a dietary supplement suppress the majority of phase I reactions, particularly those that are involved in the metabolic activation of carcinogen and raises glutathione-S-transferase (GST) activity, a phase II enzyme [37]. One of the most significant families of phase II antioxidant enzymes, GST lowers cellular oxidative levels by conjugating the substrate with reductive glutathione. Unlike neem flowers, NLE only causes a variety of antioxidant phase II enzymes in treated mice, but it has no discernible effect on phase I enzymes Glutathione S-transferase-pi (GSTPi) is expressed in tumor cell lines and human peripheral blood lymphocytes when neem extract is administered in vitro . NLE therapy has been shown to increase glutathione levels in the liver and extrahepatic organs [38]. These NLE-treated mice show a considerable decrease in the incidence of skin papillomas caused by DMBA and stomach tumors caused by benzo(a)pyrene. Both animal models also show a considerable reduction in tumor burden [88]. The DMBA- induced rat mammary carcinogenesis model [33], MNNG-induced carcinogenesis model ,and DMBA- induced HBP oral carcinogenesis model all show consistent changes in metabolic enzymes and cellular redox levels. In the liver, blood circulation, and tumor tissues of mice, NLE reduces protein oxidation, lipid oxidation, and peroxidation as markers of cellular oxidative state [38]. Interestingly, different NLE fractions exhibit varying levels of free radical scavenging and antioxidant capacity, most likely as a result of the variations in the bioactive components found in each fraction.
• NEEM COMPONENTS MODULATE TUMOR IMMUNE ENVIRONMENT :
The immune system is one of the many elements that make up the tumor microenvironment, and it may play a monitoring function. The effects of neem’s seeds and blossoms on the immune system have been somewhat investigated, despite the fact that the immunomodulating properties of the plant have been thoroughly investigated utilizing NLE. In Ehrlich cancer cells, NLE exhibits no cytotoxicity. Mice treated with NLE before being inoculated with cancer cells have a lower tumor burden and a higher survival rate [39,40]. Rather than directly affecting tumor cells, NLE inhibits tumor growth by increasing immune activity, according to another research. NLE stimulates peripheral blood mononuclear cells (PBMCs) to cause apoptosis in addition to limiting the growth of tumor cells . The surface NLE favors an active immune response, which changes the surface marker profile in PBMC from patients with head and neck cancer as well as healthy individuals . Cluster of differentiation-40 (CD40) is expressed more by monocytes in PBMC after NLE therapy, whereas CD40 ligand (CD40L) is expressed more by CD56+ lymphocytes [41]. Macrophages are stimulated to release more interleukin-12 (IL-12) when CD40–CD40L is upregulated. Natural killer (NK) cells are activated by elevated IL-2 expression because it triggers the NK cells’ perforin– granzyme B mechanism. Additionally, NLE promotes PBMC to secrete the cytotoxic cytokines tumor necrosis factor-alpha (TNF-?) and interferon-gamma (IFN-?) [42]. The supernatant obtained from these cells raises the amount of caspase while decreasing the amount of Bcl-2.Through a mechanism unrelated to elevated cytokine production, the downregulation of cyclin D1 also appears to decrease the growth inhibition of tumor cells treated with NLE . One of the most important organs in immune control is the spleen. NLE treatment has been shown to increase splenic weight in mice and to improve splenic macrophage activity, as indicated by elevated expression of the activation marker CD44 Higher amounts of IFN-? and TNF-? are secreted by spleen cells taken from mice treated with NLE . When exposed to NLE, the peripheral blood and spleen also exhibit markedly increased levels of NK cells, NK-T cells, CD4+ helper T cells, CD8+ cytotoxic T cells, and monocytes [43]. Exposure to NLE causes T cells to express the activation marker CD25 more, and monocytes to express MAC-3 more, both of which indicate macrophage differentiation [44]. Furthermore, when co-incubated in vitro, PBMC and spleen mononuclear cells obtained from these NLE-treated mice exhibit increased cytotoxicity against Ehrlich’s carcinoma cells [39]. In the Balb/c mouse model, NLE treatment reduces the tumor nodules of lung sarcoma and lymphosarcoma in the liver. This is probably because NLE-enhanced immune responses in these mice have cytotoxic effects [44]. Additionally, NLE increases vaccines’ immunogenicity and could be used as a vaccine adjuvant [47]. Since B16 melanoma cell surface antigen (B16MelSAg) is not very immunogenic, it is not recommended as a possible vaccine strategy.
Since B16 melanoma cell surface antigen (B16MelSAg) is not very immunogenic, it is not recommended as a possible vaccine strategy. According to in vivo research, the B16MelSAg vaccine’s inclusion of NLE promotes the production of B16MelSAg-targeting antibodies and causes antibody-dependent cellular cytotoxicity (ADCC) against tumor cells that express B16MelSAg . These ADCC effects have been observed in both in vitro and in vivo investigations. In mice, the growth of B16 melanoma tumors is inhibited more by the enhanced immunogeneity brought about by the vaccine-NLE combination than by the vaccine alone [47]. Breast tumor-associated antigen (BTAA), a surface antigen with minimal immunogenicity, is found exclusively in breast cancer cells and tumors, much like B16MelSAg.
• CONCLUSION AND FUTURE PROSPECTIVES :
Neem-derived chemicals are promising possibilities for anticancer therapy because of their easy availability, affordability, and human safety as a natural resource. Neem is a possible preventive and therapeutic agent against many forms of cancer, according to preclinical investigations. Neem extract’s anticancer properties are linked to the modification of key characteristic processes in tumor cells, such as the suppression of excessive growth, the induction of cell death,Different Neem components have immunomodulatory effects, which suggests that neem has unrealized potential as a therapeutic vaccine. To fully realize the enormous potential of Neem and its components in the prevention and treatment of cancer, larger clinical trials are recommended
REFERENCES
Samiksha Thorat*, Ashok jagdale, Azadirachtin Indica: A Potential Substance with Anti-Cancer Properties and Activities, Int. J. of Pharm. Sci., 2024, Vol 2, Issue 12, 566-578. https://doi.org/10.5281/zenodo.14278587
10.5281/zenodo.14278587
