Biomedicine and Chemical Sciences

Calotropis Procera (Aiton) Dryand is a soft-wooded, perennial shrub that is a member of the Asclepiadaceae subfamily of the Apocynaceae family. It has long been used in traditional medical applications in North Africa, the Middle East, South Asia, and Southeast Asia. It has been utilized for fuel, fiber, feed, and lumber since ancient times [1]. It is a lactiferous, xerophytic plant that grows naturally in arid and droughty areas. The Latin name Procera means "Presence of Wax" in the plant's shooting portion, while the Greek word Calotropis means "Beautiful," referring to the flowers. Traditional medicine has traditionally employed it [2]. It is a perennial shrub that is a member of the Apocynaceae family and is also referred to as Aak, King's Crown, Rubber Bush, Sodom Apple, and Rubber Tree. This xerophytic, evergreen, softwood plant is found all over the world, although it is particularly prevalent in the arid and semi-arid tropical and subtropical regions of Asia and Africa [3]. Aiton, Milkweed, Calotrope, Giant Milkweed, Indian Milkweed, Wild Cotton, Rubber Tree, Usha, Aak, and Madar are some of its other names [4]. Calotropis Procera usually grows to a height of 2.5–4 m in arid and semi-arid environments. This plant is of importance due to its ecological, economic, and antibacterial qualities [5]. It is distinguished by enormous pale green leaves (Figure 1) and a toxic milky sap in its green fruits. In extremely stressful situations, such as high temperatures and aridity, drought, high vapor pressure deficit, saline and wet environments, and high photosynthetic active radiation, it can withstand, adapt, and sustain growth and productivity. With a yearly rainfall of up to 150 mm, it can flourish throughout extended dry spells. Without irrigation or fertilizer, it thrives rapidly on a range of soil types in dry, desert environments [6]. The Asclepidaceae family includes the flowering plant Calotropis Procera, which is indigenous to Saudi Arabia, North Africa, Pakistan, tropical Africa, Western Asia, South Asia, Israel, and the Indo-China region [7]. It is primarily found in the wastelands of Assam, West Bengal, Rajasthan, and Punjab in India [8].

Figure: 1. Calotropis Procera.

Classification of Calotropis Procera

The Vernacular Names and Taxonomical Classification of Calotropis Procera are given below in Tables 1 and Table 2 [9,10].

Table 1: Vernacular Names of Calotropis Procera [9].

Table 2. Taxonomical Classification of Calotropis Procera [10].

Botanical Description

The bush Calotropis Procera can reach a height of five meters. Its enormous, green, obovate leaves, which are around thirty centimeters in length, are what identify it (Neuwinger, 1996) [11]. Dicot, shrub. The plant has upright stems that branch upward from the base and contain a milky white sap (Figure 2). Younger stems have a whitish-colored hair covering, a smooth texture, and a greyish-green tint. The light brown bark of mature stems is severely fissured and resembles cork. A cluster of three to fifteen flowers is formed by inflorescence axillary cymose (Figure 3). White flowers with purplish tips that are hermaphrodite, actinomorphic, free, imbricate, sepals (5), united, valvate, stamens (5), united, epipetalous, ovary superior, and two locular [12]. Fruits are huge, sub-globose, greyish-green follicles with many brown, flattened seeds and a tuft of long, white, silky hairs on the axile placenta [13]. Simple leaves (Figure 4), on the other hand, have a milky white sap, are oval in shape, sessile, have an acute apex, and have a glabrous upper surface. The underside may have a tuft of stiff hairs (Figure 5) at the base of the midrib or be heavily coated in small white hairs [14].

Chemical Constituents

All parts of the plant release white latex when they are cut or broken, which serves as a barrier against viruses, fungus, and insects. A and B amyrin, giganteol, gigantin, teraxasterol, and b-sitosterol isogiganteol are present throughout the entire plant. Numerous flavonoids, triterpenes, cardiac glycosides, and sterols are among the many secondary metabolites that this plant has generated [15]. The cardiac glycosides calotopin, uscharin, calotoxin, calactin, uscharidin, and gigantin contained in this plant's white, milky latex have been shown to have significant wound-healing properties [16]. Numerous chemicals, including cardenolide, triterpenoids, alkaloids, resins, anthocyanins and proteolytic enzymes in latex, flavonoids, tannins, sterol, saponins, and cardiac glycosides, have been identified through phytochemical research on Calotropis [17]. However, Calotropis gigantea leaves contain five main chemicals: Pleurone, Calotropagenin, Calotoxin, (+)-dehydrovomifoliol, and Methyl β-carboline-1-carboxylate [18]. Numerous compounds, including carenolide, triterpinoids, alkaloids, resins, anthocyanins, and proteolytic enzymes in the latex, have been identified by studies on the phytochemistry of Calotropis. Flowers contain terpenes, cyclisadol, and multiflorenol. The main substances included in the leaves are amyrin, amyrin acetate, -sitosterol, urosolic acid, cardenolides, calotropin, and calotropagenin [19].

Phytochemical Screening

The method of qualitatively identifying secondary metabolite chemicals in plants is known as phytochemical screening [20]. The first step in an extraction procedure is choosing a suitable solvent, such as water, ethanol, acetone, ethyl acetate, n-hexane, and other organic solvents [21, 22]. The separation of active compounds from a plant, such as polar, semi-polar, or non-polar secondary metabolite compounds like alkaloids, flavonoids, tannins, saponins, terpenoids, and phenols, is significantly influenced by the polarity of a solvent [23]. Variations in C. gigantea's secondary metabolite composition can be attributed to genetics, growth circumstances, and geography [24]. According to a number of research, the results of phytochemical screening of C. gigantea plant extracts vary depending on the different regions of Aceh province, including geothermal sites [25], mountainous [26], and coastal areas [27].

Pharmacological Properties

Because of their pharmacological and therapeutic qualities, plants have been utilized for ages as a source of natural treatments [28]. Calotropis Procera's pharmacological qualities were once widely employed to cure a variety of human ailments, such as fever, leprosy, asthma, rheumatism, dermatitis, dysentery, indigestion, diarrhea, and elephantiasis. Alkaloids, flavonoids, steroids, tannins, terpenoids, saponins, and cardiac glycosides are among the metabolites found in many plant sections that have been identified by numerous investigations as potential materials for the green synthesis of metal ions [29]. The dried leaves of Calotropis Procera are used in Ayurveda as an expectorant and to relieve rheumatic pain and paralysis [30, 31]. In particular, migraines are treated with the delicate leaves [31]. These leaves' powdered form is also used to improve wound healing, act as a laxative, and treat indigestion. A decoction prepared from the aerial portions of Calotropis Procera is used in traditional Saudi Arabian medicine to treat fever, joint pain, constipation, and muscle soreness [32]. Additionally, Calotropis Procera has shown promise in treating sinus fistula, diarrhea, and a variety of skin conditions [33]. Its root bark is used in India to treat intestinal worms, skin conditions, and enlargements of the abdominal viscera. Milky latex is used to treat skin conditions in Senegal [34].

Anti-Cancer Activity

Hexane, dichloromethane, ethyl acetate, acetone, and methanol extract of Calotropis Procera stem [36, 37], as well as aqueous extracts of dry latex [35]. have been described as having anticancer properties. The DL of Calotropis Procera was found to offer complete protection against the formation of hepatocarcinogenesis when tested in a transgenic hepatocellular cancer model in mice. While Huh-7 and COS-1 cells had severe cell death and a marked drop in serum vascular endothelial growth factor levels, AML12 cells were found to be alive. Calotropis gigantea's cardiac glycosides have the ability to stop the growth of cancer and tumor cells. In vitro, the expression of p53 and Bcl-2 genes in breast cancer CF-7 can be influenced by cardiac glycosides like calactin, calotropin, asceplin, and cymarin. It is more plausible that Bcl-2 regulation, which disrupts antiapoptotic activity, was responsible for the tumor suppression actions on the mediated test sample than p53, which promotes apoptosis [38].

Antifertility

Albino rats were utilized in the study to examine the hormonal and antifertility effects of an ethanolic extract of Calotropis Procera roots. Significant anti-implantation (100% inhibition) and uterotropic action were observed at a dosage of 250 mg/kg (1/4 of LD50). Antiestrogenic effect was not demonstrated [Saxena and Saxena, 1979] [39].

Larvicidal Activity

Calotropis procera showed high larvicidal effect when tested against Anopheles labranchiae mosquito larvae, with an LC50 (24 hours) ranging from 28 to 325 ppm. The detrimental effects of crude extracts of Calotropis Procera (from both leaves and flowers) on the termite species Heterotermes indicola and Coptoter mesheimi have been studied [40]. The ethanolic extract of Calotropis gigantea has been shown in earlier studies to have larvicidal effects on Ae. aegypti larvae, with an LD50 value of 351.43 ppm. Calotropis gigantea and Calotropis procera had similar effects on Ae, as evidenced by the LC50 values of all sections for the larvicidal action. larvae of Aegypti [41]. Calcotoxin and calcotropin are the main compounds that may be responsible for the larvicidal characteristics of the Calotropis species.

Analgesic Activity

The dry latex (DL) of Calotropis Procera has been shown to have analgesic qualities [42]. A dose of 415 mg/kg of DL was more effective in suppressing writhing brought on by acetic acid than an oral dosage of aspirin (100). The alcoholic extract of Calotropis gigantea flowers has analgesic properties. An oral dosage of this extract generated a notable result in the activity, which was conducted using the writhing test caused by acetic acid and the hot plate technique. The effects of an alcoholic peeling extract of Calotropis gigantea roots (at oral dosages of 250 and 500 mg/kg body weight) on the central nervous system (CNS) of albino rats were investigated. Both the writhings caused by acetic acid and Eddy's hot plate approach demonstrated significant analgesic advantages [43].

Antiviral Activity

To measure antiviral efficacy, two-fold serial dilutions of both RFL (rubber free latex) and SLP (soluble laticifer proteins) ranging from 10 mg/mL to 0.019 mg/mL were mixed with a standard quantity of the virus and incubated for 30 minutes at 37 °C. 22 Nine-day-old chicken embryos from Big Bird Laboratories in Lahore were injected with the mixture via an allantoic cavity route under sterile settings. The negative control was created by injecting PBS (phosphate-buffered saline) into the embryo, whereas the positive control was created by injecting the virus alone. Every embryo was incubated at 60% humidity and 37 °C. Every day, an egg candler was used to assess the embryos' viability. The spot hem agglutination approach was used to detect the presence of virus in chorioallantoic fluid that was collected in sterile test tubes [44].

Allelopathic And Insecticidal Potential

The growth of Brassica oleracea var. botrytis (broccoli), suppression of some plant species, and allelopathic potential of Calotropis Procera (giant milk weed) have all been observed [45, 46]. Additionally, C. procera's allelopathic activity was evaluated against a variety of weeds, including Portulaca oleracea (pigweed), Dactyloctenium aegyptium (Crowfoot grass), Bidens pilosa (Spanish needle), and Chenopodium murale (salt green) [47]. Aqueous extracts from Calotropis Procera shown insecticidal efficacy against Henosepilachna elaterii Rossi, the melon lady bird beetle [48].

Assessment of the Antimicrobial Activity of Calotropis Procera Using Agar Well Diffusion

The agar well diffusion method was used to assess the antibacterial activity of plant extracts from Calotropis Procera leaves. After the test organisms were inoculated on Muller Hinton agar and given two minutes, five 5.0 mm wells were drilled into the agar plates using a sterile corked borer. Four of the wells received 0.3 mL of various plant extract concentrations (200, 100, 50, and 25 mg/mL). For a whole day, agar plates were incubated at 37°C. The test for the fungi was conducted on SDA plates and incubated for 72 hours at 30 degrees Celsius. Following incubation, the diameter of the inhibitory zones was measured and recorded to the closest millimeter [49].

In Vitro Anti-Sporozoite Activity of Calotropis Procera

The length of the incubation period and the extract concentration rate were used to measure the sporozoite vitality rate in Calotropis Procera. When compared to the control treatment (K2Cr2O7), 1,000 µg/mL of extract and 25 µg/mL toltrazuril were found to exhibit statistically significant alterations (p <.05) in the viability rate of E. magna sporozoites after 12 and 24 hours of incubation. However, at lower extract concentrations (500, 250, and 125 µg/mL), the sporozoites showed different levels of vitality (Figure 6) [50].

Figure: 6. The Results of the Effects of Calotropis Procera Leaf Extract In Vitro

Vitality on E. magna Sporozoites after 12 and 24 hours. Note: * significant differences compared to the control and 25 µl/mL of toltrazuril (p < 0.05, n = 3).

Antifungal Activity

With differing degrees of growth inhibition, the aqueous leaf and root extracts of Calotropis Procera showed significant antifungal efficacy against A. niger and R. stolonifer. The fungal isolates' mycelia growth was suppressed by both leaf and root extracts, although the leaf extract was more effective, suppressing 81.1% and 79.4% of A. niger and R. stolonifer, respectively. These results are in line with earlier research by Hassan et al. [51] and Aliyu et al. [52], which also demonstrated Calotropis Procera's strong antifungal qualities against a variety of fungal species and supported its potential as a fungistatic or fungicidal agent at comparatively low concentrations.

Antimicrobial Potential Against Fungal and Bacterial

Researchers found that the Calotropis Procera plant extract was effective against a few pathogenic bacteria [53–59]. Calotropis Procera leaf extract at different doses (1%, 2.5%, 4%, 5.5%, and 7%) shown antifungal potential against Macrophomina phaseolina (Tassi) Goid on mung beans, a significant pulse crop [60]. The antibacterial properties of the Calotropis Procera essential oil therapy were tested against two fungal and seven bacterial species. Similarly, when Calotropis Procera leaf extract was present, the iron Phyto-nanoparticles demonstrated their antifungal ability against Alternaria alternata [61]. Calotropis Procera roots contain bioactive chemicals that were used to treat infections. Calotropis Procera latex has been shown to have anthelmintic properties against Haemonchus contortus infection in Najdi sheep [62]. White rot fungi were identified by Aswal et al. [63] in order to break down the lignin in Calotropis Procera fiber.

Anthelmintic Activity

The in vitro and in vivo anthelmintic activity of the Calotropis Procera flower extract was compared to that of levamisole. The crude aqueous ex tract (CAE) and crude methanolic extract (CME) of flowers had a substantial impact (P<0.05) on live Hemonchus, according to the in-vitro investigation. For an in-vivo investigation, sheep that were naturally infected with gastrointestinal nematodes were given the crude powder (CA) and extract. At 3000 mg/kg, the proportion of eggs in the sheep treated with CAE and CP decreased by 88.4 and 77.8%, respectively, on days 7 and 10. However, lambs treated with CME only displayed a 20.9% decrease in egg count. Based on the aforementioned results, it was determined that the plant extract had notable activity against nematodes, whereas levamisole's activity was 97.8–100% [64].

Wound Healing Activity

The body goes through a healing process called wound healing when the skin or other soft tissues are damaged. The underlying cells experience an inflammatory reaction and increase the creation of collagen when the skin is damaged. New epithelial cells develop over time. A sterile latex solution (20 µL) was administered topically once day for seven days. The latex stimulated the creation of collagen, DNA, and proteins as well as epithelization, which decreased the size of the wound [65]. After the extract was administered, the epithelialization time decreased statistically significantly (p<0.001) in both normal and dexamethasone-treated rats. This reduced the task's completion time from 28 days to 17 or 18 days. Rats treated with dexamethasone showed comparable increases in breaking strength after consuming the same extract [66]. The wounds in the honey and triamcinolone groups with 50% latex were much smaller after 7 days, and they were even smaller after 14 and 21 days [67]. Triterpenoids -amyrin, flavonoids, cardiac glycosides, cardenolide anthocyanins, mudarine, lupeol, -sitosterol, flavanols, resin, a powerful bacteriolytic enzyme called calactin, a nontoxic proteolytic enzyme called calotropin, and a wax are all found in the plant, according to its phytochemistry [68]. The extract significantly increased strength upon breaking. Additionally, the rate of wound contraction was significantly increased, and wounds treated with extract underwent faster epithelialization.

Anti-Hyperglycemic Effect

In the current study, found the antioxidant and anti-hyperglycemic properties of Calotropis Procera dry latex (DL), which has strong anti-inflammatory action, against alloxen-induced diabetes in rats. Hepatic glycogen content increased and blood glucose decreased in a dose-dependent manner when DL was taken orally every day at doses of 100 and 400 mg kg-1. DL's anti-diabetic and antioxidant properties were on par with glibenclamide, a common anti-diabetic medication [69].

Anti-Malarial Activity

The IC50 values of the ethanolic extracts of the various Calotropis Procera parts against P. falciparum MRC20-CQ-sensitive ranged from 0.11 to 0.47 mg/ml. Additionally, flower and bud extracts were the most effective against MRC76_CQ-resistant bacteria, ranging from 0.52 to 1.22 mg/ml. These extracts are 220–440 times less effective than CQ, yet they nonetheless merit more research to identify the key ingredients. The ethnobotanical use of this plant is supported by the results [70].

Anti-Diabetic Activity

The antidiabetic properties of compounds extracted from various plant sections can be investigated. A review paper about Calotropis Procera's components and pharmacological characteristics shed light on a few of the plant's isolated chemicals. It has been claimed that various portions of the plant contain compounds like calotropin, calotropagenin, isorhamnetin-3-O-rutinoside, calotoxin, calactin, uscharin, and others [71]. Numerous in silico investigations discovered inhibitors from a wide range of natural compounds that target important receptors in the disease mechanisms, such as alpha glucosidase [72–78]. Finding potentially novel AGIs from plants is a crucial foundation for drug discovery since inhibition of these enzymes is directly linked to the treatment of diabetic mellitus [79–84].

Miscellaneous Activities

Sayed et al. (2016) investigated the antiapoptotic activity of Calotropis Procera latex on catfish exposed to 4-nonylphenol (100 μg L−1) as a chemical pollution. The antiapoptotic effect of the crude latex against the toxicity of 4-nonylphenol was confirmed by a significant (P<0.05) decrease in apoptotic cells, ions, and enzymes (Superoxidase dismutase, acetylcholinesterase, cortisol, etc.). Therefore, 4-nonylphenol's toxicity was countered by the antiapoptotic effects of crude latex. Phenylhydrazine and paracetamol-induced Wistar rats were used to assess the anti-hyperbilirubinemic effect of leaves. The bilirubin-lowering effect of Calotropis Procera aqueous extracts was demonstrated by a significant (P<0.05) drop in serum total bilirubin concentrations in hyperbilirubinemic rats. Because Calotropis Procera contains bioactive phytochemicals with therapeutic potential, recent research has shown that it has a much wider variety of positive effects. Basic research has demonstrated additional actions, but only cytotoxic tests on cancer cell lines have been thoroughly established in clinical trials. The majority of research is restricted to in vitro investigations that do not examine the molecular mechanism of action. In order to understand the underlying mechanism associated with traditional usage, mechanism-based in vitro and in vivo research should be conducted [85].

Toxicological Profile

Calotropis Procera's toxicity has been extensively studied in a variety of biological systems. Despite having a number of pharmacologically active ingredients, it has been shown in multiple trials to have harmful consequences, particularly when used in non-standardized doses or improperly prepared. The heatmap for the toxic compounds of Calotropis Procera Shrub [86] and acute and chronic toxicity, cytotoxicity, genotoxicity, neurotoxicity, and dermal and ocular risks are all included in the toxicological profile (Dogara, 2023).

The Commercial Use of Drugs

Drug resistance for the treatment of infectious diseases in human pathogenic microorganisms has developed as a result of the commercial usage of medications. Furthermore, antibiotics are seldom linked to a number of adverse effects, including hypersensitivity, immunological suppression, and allergic reactions. Prehistoric use of natural goods, including as herbal fractions and isolation of anti-microbes, has been shown to be safe and beneficial in a number of ancient systems. The most recent advancement in the usage of herbal antimicrobials has grown and demonstrated their significance. Tumors and other swellings of the neck are treated externally using the boiling residue of the Calotropis Procera plant, particularly paste. Plant aqueous extract is a common cancer treatment in the ethnic medical system [87].

Industrial And Novel Applications

Calotropis Procera has drawn interest for a number of industrial and biotechnological uses in addition to its therapeutic value. Strong, lightweight, and moisture-resistant fibers from the plant's stem bark can be used to make ropes, carpets, fishing nets, and environmentally friendly textiles [88]. Rubber-like substances and proteolytic enzymes found in Calotropis Procera latex have been investigated for use in biodegradable materials, bioadhesives, and enzyme-based businesses. Additionally, because of their capacity to effectively break down proteins, its proteases have demonstrated promise in waste management and leather processing [89]. The plant's function in the green synthesis of nanoparticles, where extracts are employed as stabilizing and reducing agents for the creation of metal nanoparticles like silver and gold, has also been highlighted in recent studies. These nanoparticles show improved antibacterial and anticancer properties, suggesting potential uses in drug delivery systems and nanomedicine [90].

Traditional Uses

India is a well-known tropical nation rich in natural resources and traditional knowledge about how to harness them. One such important medicinal plant that is highly prized for its numerous ethnobotanicals uses and huge commercial significance is Calotropis Procera [91]. Certain plant parts have historically been used medicinally to treat a variety of illnesses. Rheumatism, fever, indigestion, colds, eczema, diarrhea, boils, and jaundice have all been treated with the entire plant. Eczema, leprosy, elephantiasis, asthma, cough, rheumatism, and diarrhea have all been treated with the root; leukoderma, intestinal worms, and leprosy have all been treated with the stem [92,93]. Due to its antifungal qualities, latex is used in drugs to treat tinea capitis in children. However, there is a history of more extreme uses of latex that cause abortions in women [94]. The blossoms were believed to improve appetite, reduce mucus accumulation, and improve digestion. The blossoming tips were used to treat asthma. Additionally, the root bark was used to treat elephantiasis [95,96].

Conclusions and Future Prospects

Calotropis Procera is a plant with a variety of biological traits that make it both a potentially invasive species and a significant therapeutic and socioeconomic species. The goal of this discussion is to evaluate its growing worldwide spread, important biological and ecological characteristics, applications in both conventional and cutting-edge fields, and infestation as an environmental weed. Additionally, it is an effort to identify the areas of ongoing study that have received less attention and knowledge gaps.

The plant's general biological and ecological characteristics-especially those pertaining to adaptations or plasticity-have not been thoroughly studied, despite the fact that its medicinal and industrial uses have received the attention they deserve. Furthermore, there hasn't been much attention paid to Calotropis Procera's toxicity-bioactivity relationship, which is crucial for confirming its therapeutic characteristics. Assessing these fundamental aspects could enhance its economic use and open the door to other uses. In addition, filling in these knowledge gaps can aid in comprehending its invasive behaviour and any future dangers to the environment or biodiversity.

In order to control or contain Calotropis Procera in a timely manner, it is also necessary to map its existing and potential spread. Mechanical, chemical, or biological techniques can be used to effectively inhibit the spread of Calotropis Procera in the invaded ranges. This can be followed by ongoing monitoring over the following few years to prevent the emergence of new plantlets. Acknowledging the plant as a significant environmental weed can enhance scientific, legislative, stakeholder, and municipal control initiatives. Additionally, encouraging its use on both commercial and non-commercial levels may be a financially feasible, or more accurately, profitable, approach to its management.

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