Review Article pepo and in the Management of Anti-proliferation by JAK/STAT Pathway

Souvik Mukherjee, Dilipkumar Pal* Department of Pharmaceutical Sciences, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur, Chhattisgarh, INDIA.

ABSTRACT () is capaciously recycled similar to food and in folk medicine

throughout the world. It accords to genus Cucurbita (CCT) under family . There are a plenty of important medicinal phyto-constituents belonging to cucurbitoside AR like triterpenoids, C T and CCT . A survey of the literature demonstrates that C. pepo, has the capacity to improve prostatic hyperplasia, urinary dysfunction and cytotoxic properties. Many pharmacological revisions have established its role in hepatoprotection, st n G st inhibition of Pr gland cancer (CNCR), anti (A t) oxidant effects, inhibition of Lu , BR and st triple-negative BR CNCR by blocking JAK/STAT signaling (Sgls) pathway (Ptw). It has also n n n n A t microbial, A t -inflammatory, A t -diabetic and A t ulcer activities by supporting its

traditional claims. Establishment of C. pepo and cucurbitacin (CCBT) in the management n of A t -proliferation by JAK/STAT Ptw. Data towards writing this review are generated through exploration of different websites like MEDLINE (PubMed), Google Scholar, Science Direct, Scopus, Cochrane, SID and Magiran databases. We have selected 2016-

2018 duration for the same purpose. We have found 88 papers related to this topic. CCBT

is found to arrest unlimited cell (CEL) division and respective apoptosis (Appt) in vitro and

in vivo CNCR models. A plenty of molecular design targeting CCBT have been invented, such

as fibrous-actin, glsS transducer and activator of transcription (STAT), cyclooxygenase-2,

etc. This review is minded at CCBT from C. pepo which dwindle the proliferation of human

CNCR CEL through the JAK/STAT Ptw. Key words: Anticancer activity, Cucurbita pepo, Cucurbitaceae family, JAK/STAT pathway, Cucurbitacin, Cyclooxygenase-2.

Submission Date: 10-01-2020; Revision Date: 17-07-2020; INTRODUCTION Accepted Date: 20-01-2021

CNCR lies its uniqueness to the maximum have restrictions subsequent from the DOI: 10.5530/ijper.55.1.3 Correspondence: normally identified diseases SEAS (D ) and is expansion of resistance to the treatment. Dr. Dilipkumar Pal associated with ill health and death set up The identification of defensive molecules Associate Professor, causing a health problem globally. Even starved of side effects ruins a crucial Department of Pharmaceutical Sciences, though unlimited determinations have been independent in the fight against CNCR. The Guru Ghasidas, Vishwavidyalaya, found ready to find out a remedy, CNCR additional choices goal next to the initial Bilaspur-495 009, remnants a very projecting cause of death in finding of CNCR in the preliminary stage can Chhattisgarh, INDIA. Phone: +91 077522-258878 humans. Carcinogenesis (CCNG) is a different assist with its appropriate supervision. In E-mail: [email protected] step and different factorial process including the meantime, plant (PLt)-derived products the incidence of vibrant and disconnected have taken a major role to inhibit numerous molecular and CEL modifications. There are chronic DSEAS, as well as CNCR. The use of PLt different but thoroughly associated stages substances to inhibit or defer the growth of of origination, elevation and development CCNG has been called for chemoprevention 1 are found in CNCR. Present-day CNCR and there is a rapid increasing attention treatments, chemotherapy, targeted agents, towards the usage of natural compounds as radiation, surgery and immunosuppression probable chemo-protective and therapeutic www.ijper.org

Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 1 | Jan-Mar, 2021 1 Mukherjee and Pal.: Cucurbitacin as a JAK/STAT Pathway Inhibitor

2 agents. Pumpkin (PMPN) seed (SEd) has several health of PMPN consist of the United States, Canada, Mexico, 10 benefits. MPNP is refined all over the biosphere for usage India and China. as root vegetables as well as medicine.3 It is also recycled Plant Characteristics by tradition as medicine in many nation-states such as China, Yugoslavia, Argentina, India, Mexico, Brazil PMPN is yearly parsley with heavy mounting stems. The root is thriving established and towards 40 cm and America. Its extensively detained medicinal (Medc) usages have concentrated investigation with modern unfathomable with 5m extended. The stems are a branch n n off, enclosed in spongy white up to 10 m long and implements and recognized with good A t -diabetic, A t n m frequently yield extrinsic roots at nodes. The petioles are -hypertension, A t –tumor(t R), immunomodulation, n n n 5-20 cm long. The tinny leaves are alternative, modest, A t bacterial, A t -hypercholesterolemia, intestinal A t -parasitic, An -inflammatory and analgesic properties.4 palmate, veined, round to reniform, essentially cordate, t apically obtuse, unsubdivided to trivial 5-7 lobed, Cucurbita pepo 7-30 cm across, wide-ranging than long, stark to soft blooming and finely margin with toothlike projection, PMPN (Cucurbita pepo) is one of the eldest identified 3-5 rounded or obtuse, apiculate lobules, the central one nurtured classes of shrub. It accords to the genus CCT bigger than lateral ones.11,12 Unisexual flowers of P and family Cucurbitaceae or CCT and contain crops like MPN . Ethno-pharmacological studies display are aromatic. The calyx is enclosed in white pubescence that C. pepo is recycled in various countries for treating and bears 5 free sepals, 0.5-2 cm long. The corolla is yellow to orange color, tubular at least 5 cm long and many DSEAS like , viral infections, pain, urinary disorders, ulcer, diabetes and oxidation.5 Mainly broad. Staminate flowers are about 10-23 cm long. Pistillate flowers are grown on shorter pedicles, only up Ayurveda system has used to assess segment of the PLt as well as corpuscles of the fruits ( ut) and S . The S to 5.5 cm long and have an inferior 1- ocular ovoid ovary Fr Ed Ed ut are recycled to treat the problems of urinary system, with a short thick style with 3-5 bilobed stigmas. Fr are 13 st are highly inconstant in shape, color and size. The hypertension, kidney stones, prostate (Pr ) DSEAS, Sk shape is elongated, cylindrical, oval, flattened, globular, erysipelas skin ( N) infection and carcinomas (CArom). Exact cultivars of winter squash resulting from other heart-shaped and tapering to a curved neck on one or species such as C. argyrosperma and C. moschata, are also both ends. The length is from 5.8 to 71.6 cm and width Sk 6 at times called “ P . from 11.2 to 48.6 cm. The N can be smooth, wrinkled. MPN The flesh, variable in color and thickness can be white, Taxonomical Classification ofC. pepo yellow, or orange and 1 to 6.4 cm thick14 [Figure 1]. Taxonomic classification.7 S Characteristics Kingdom: Plantae Ed P S is also recognized as pepita. The S are Subkingdom: Tracheobionta MPN Ed Ed characteristically flat, unequally oval, green in color Super division: Spermatophyta and typically enclosed by a white husk [Figure 2].15 P Division: Magnoliopsida MPN SEd produce 34-54% oil. The size and weight of the Subclass: Dileniidae ut SEd rise as the Fr size rises, lining up between 1.6 to Order: Violales 2.9 cm long, 0.7 to 1.6 cm wide and 0.28 to 0.69 cm

Family: Cucurbitaceae thick. The SEd seem doable for 6-8 years but abide by no

Genus: CCT L endosperm and the embryo embody leaf-like cotyledons and a short radicle.16 P S oil is popular as succulent Species: CCT pepo L MPN Ed oil and also used as a nutritious food. PMPN SEd and its Vernacular Names oil are prosperous in phytosterols, polyunsaturated fatty Hindi: Safed Kaddu, Kumrha Marathi: Kohala, Bhopli n AR acids (FTA), A t -oxidant vitamins, carotenoids (C T), Telugu: budadegummadi, Bengali: Safed Kaddu, Tocopherols and its versatile facets such as protein, Sanskrit: karkaru, kurkaru and kurlaru, kushmanda magnesium, copper and zinc. Due to the presence 8,9 English: squash. of these constituents, PMPN are recognized as a good reservoir for providing many health remunerations.17 Habitat Chemical composition of C. pepo PMPN are full-fledged throughout the biosphere for a diversity of explanations extending from agronomic These are categorized through a low contented fat ulterior motive. Only Antarctica is not capable to (2.3%), mono-di-poly saccharides (66%), proteinoids AR harvest PMPN. The major international manufacturers substances (3%) and high C T contented with

2 Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 1 | Jan-Mar, 2021 Mukherjee and Pal.: Cucurbitacin as a JAK/STAT Pathway Inhibitor

18 t 26-28 magnitudes of 171.9 to 461.9 microgram. The mineral β-crypto X n are also present. Acylated phenolic ly investigation specified that MPNP pulp is enclosed with glycosides(G o) such as cucurbitoside F, H, I, K, L, M, 19-21 CR CR great levels of elements which shown inTable 1. The 23-24- dihydro CCBT lariciresinol (L S), seco-iso L S, iso structure of P S is reasonably varying. The content CR CR ly CR ly MPN Ed L S, L S -4’-o-β-D- G o, L S - 4-o-βeta-d G o, (24s)- C of amino acids (A d), FTA and minerals may differ stigmata- 7,22E, 25-trien- 3-one, (24s)-stigmasta-7,22 E, significantly, depending on changed conditions. Such 29-31 25 –trien-3beta-ol, CCBT L 2-O-β-D-glucopyranoside. changes may be affected by differences in cultivar or Others Phytoconstituents are also exhibited in Table 3. origin. PMPN SEd contain 50% fatty oil which is dark green Anti- CNCR Mechanism of C. pepo and rich in free FTA. The arrangement of FTA differs on CT numerous factors(F ) like the variety of places where CNCR is the deregulation product of programmed CEL CT the PLt are developed, weather, growth F and favoring death. Numerous favorable goals for mediation is ripeness.22 The instabilities in the oil constituents is very recognized by reviewing the molecular defects such n high, subsequent from a wide G e variation, farming as the signal transduction Ptw that control Appt. In atmosphere, storage time and storage conditions. The this viewpoint, PMPN SEd comprising of CCBT and its glyceride part content variates from 73.1% to 80.7 % derivatives have developed a new emphasis for CNCR C unsaturated FTA, mainly oleic acids (OA d) and linoleic (L drug discovery because of its durable ability to inhibit AC ). Again, the same fraction contains 19% saturated d different types of CNCR. CCBT and its byproducts inhibit F consisting of mainly palmitic (P AC ) and stearic TA d CNCR development by a comprehensive variety of acids(S AC ) (6%). Several studies have reported similar d mechanisms (Mehs), comprising of pro- Appt, installment types of data regarding proportions of total F or free TA of autophagy, CEL cycle seizure, inhibition of CNCR F in the cake fraction of P S : 29.9% L AC and TA MPN Ed d entrenchment and shifting. CCBT also modifies numerous OAC 50.4% [Table 2]. d intracellular Sgls Ptw [Figure 3]. Sgls transducers and PMPN SEd are enclosed comparatively huge quantities of K (5,790 μg/g dry weight) and chromium (approximately Table 2: List of F in P S . 3 μg/g dry weight), Na content of same S is low (6.9 TA MPN Ed Ed Name Structure Content (%) μg/g dry weight). Other minerals present in PMPN SEd are: P (15,700); Ca (346); iron (106); Mn (49.3); Al (9.21); Ba O

(1.16); Co (0.29); strontium (1.83); Ni (0.53); As (0.45) P AC 10.68+/-0.42 (in μg/g dry weight). Notable is the low amounts of d OH calcium in the SEd. One hundred-gram roasted PMPN S contain 25.94 mg Ca, 955.81 mg P and 8.06 mg of Ed O 23-25 Fe. Palmitoleic 0.58+/-0.14 AR t t C A d Numerous constituents such as C T, as lutein(L n), L n OH t t t epoxide, 15- cis- L n, 9(9’) –cis- L n, 13(13’)- cis – L n, α – CAR , β- CAR , violaxanthin(Xt ), auro Xt epimers, O T T n n C t t t t S A d 8.67+/-0.27 flavo X n, lute X n, chrysanthema X n, α-crypto X n, OH

O Table 1: List of minerals in PMPN SEd. C Components Nutrient value % of RDA OA d 38.42+/-0.37

Na 7 mg 0.5 OH

K 809 mg 17 O

C Ca 46 mg 4.5 L A d 39.84+/-0.08 OH Cu 1.343 mg 159 Fe 8.82 mg 110 O L AC 0.68+/-0.14 Mg 592 mg 148 n d OH Mn 4.543 mg 198 P 1.233 mg 176 O C Se 9.4µg 17 Ga A d 1.14+/-0.00 Zn 7.81 mg 71 OH

Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 1 | Jan-Mar, 2021 3 Mukherjee and Pal.: Cucurbitacin as a JAK/STAT Pathway Inhibitor

Table 3: List of Phytoconstituents present in PMPN SEd. Name Category Structure t AR α-crypto X n C T OH

t AR HO L n C T

HO OH

O

t AR OH L n epoxide C T Stigmastatrienol Steroid

H HO H H OH

t AR 15-cis- L n C T R Squalene T P HO

OH OH

t AR HO OH 9(9’)-cis- L n C T OH O O HO Vicine Alkaloid O NH2

N NH

NH AR 2 α-carotene C T

OH

O Viola Xt CAR OH R n T HO Kuguacin T P O H H

HO O

HO t AR O Auro X n C T O OH H

R CCBT T P OH Flavo Xt CAR n T O

HO O HO OH OHO t AR O Luteo X n C T HO O R H OH CCBT A T P HO O O H O

O Chrysanthem OH AR HO t C T X O n O O HO R H H OH CCBT B T P HO O

O

4 Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 1 | Jan-Mar, 2021 Mukherjee and Pal.: Cucurbitacin as a JAK/STAT Pathway Inhibitor

O HO O O C E TR H H OH CBT P HO O

O

O OH HO O R H H OH OH CCBT G T P HO

O

Figure 1: Pumpkin Flowers and Fruits. O HO O C F TR H H OH OH CBT P HO

HO

O HO O C M TR H H OH OH CBT P HO

O

O OH HO Figure 2: Germinated . O C K TR OH OH CBT P HO

O

OH

C L TR O CBT P O O O O OH HO O N HO OH HO

SC activators of transcription (TR ) 3 and Janus enzyme Sgls Figure 3: JAK/STAT and WNT Signaling Pathway. Ptw are the key Mehs for CCBT to speak into necrobiosis AG to place forth their compelling malignant (M L) n neoplasm impact. The capability of CCBT to prevail CEL Ptw. Protein empirical G e, enacting the obliterator of cycling in the G2/M part by diversified controllers is C communication super molecule family, binding to 32 tK additionally a major approach to fight different CNCR. JAK, represent the major negative regulators of the STAT3 (S 3) controls the exposition of genes (Gn ) 3 3 ta e JAK/ Sta Sgls Ptw. In recent past, it is accepted that Sta which intercede multiplication (e.g., c-myc and cyclin may also be called up by several alternative CtK, like IL-7, n D1), lowers activities of pro-apoptotic G e (e.g., Bcl-xL, IL-10, IL-20, leptin, WBC colony-stimulating issue and Bcl -2 and surviving) and/or accelerates maturation 33 n cuticular protein. Admist the seven human STAT G e, 3 through vascular epithelial tissue protein (VEGF). Sta , a typical oncogenic communication Ptw, is integrally

Conversely, (CtK) will inhibit the STAT-3 Sgls called up in many sorts of CNCR, together with eighty-

Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 1 | Jan-Mar, 2021 5 Mukherjee and Pal.: Cucurbitacin as a JAK/STAT Pathway Inhibitor

st n m two of glandular CArom, seventieth of breast (BR ) CNCR, A t - t R genic activity within the absence of activated 3 41 over eighty two of the CArom of the top and neck, seventy Sta . AG pls one of cavity M L neoplastic(NE ) DSEAS, over five Induction of Appt hundredth respiratory organ CNCR and five hundredth of 34 HCC, lymphomas and myelomas. Uncontrolled doings CCBT B, D, E, I and IIa influence Appt in different classes 3 m of C C by arresting the S 3 P . S 3 is a T SC FCT of Sta is unquestionable in an exceedingly form of t R NCR EL ta tw ta R st AG pls st that rolls Gn expression via cross-talk with another T SC varieties, together with BR M L NE DSEAS , Pr CNCR, e R 35,36 FCT, such as β- (Β n), hypoxia-inducible factor-1, nuclear melanoma, multiple myeloma and CNCR of the blood. c n factor-B, c-myc, c-jun and closing off S 3 stimulation Numerous G e mutations cause organic activation ta 3 influenced Appt. CCBT B has distinct structural ups and of Sta , as for example over-phrasing and organic 37 3 downs as symbol for Appt, which consists of nuclear triggering of cuticular protein receptor (EGFR). Sta m fragmentation, chromatin contraction and embodiment takes part to t R growth by widening the CEL cycle by n of apoptotic bodies. CCBT B may be significant for warding off necrobiosis and speculating onco G e like 3 c-Myc and Bcl-X. S has lately been incontestable to both inflecting the empathy of CNCR CEL to cytotoxic ta n st enhance glandular C metastasis by nurturing P lymphocyte and inspiring A t CNCR immunity by the Arom r 3 n m C C exodus. C are identified as A t agents prohibition of the JAK2/ Sta Ptw. These Mehs pinpoint NCR EL CBT t R 3 3 that prohibition of the JAK/ Sta Ptw with CCBT B may associated with other Mehs, like conflicting with Sta Sgls. They also influence the virtue of the actin cytoskeleton. be impressive in CNCR immunotherapy. Moreover, in human colon adeno CArom, the CCBT B-influenced- Appt As for instance, CCBT E wards off the propagation of glandular C C and disrupts the body architecture is sustained by a reactive oxygen species system instead Arom EL 3 38 of simple protein and supplements. However, C A, of that of Sta . CCBT D stimulates the apoptotic Ptw by CBT 3 st S 3 annihilating Sta activity in BR CNCR CEL and splitting B, E, I impede the phosphorylation (PH R) of Sta and/ or JAK2 and same way rules out S 3 deoxyribonucleic fragments to procaspase-3, procaspase -9 and PARP in ta 42-45 C 3 SC human endometrial as well as ovarian CNCR CEL. A d-attachment and Sta -mediated cistron TR in CArom 39 S A549 line. Similiarly, CCBT I causes debasement of PH R Induction of Autophagy - S 3 in the B st, P st and exocrine gland MAG N pls D ta R r L E SEAS C , specifically C B and I, activate autophagosome C lines (MDA-MB-231, MDA-MB-468 and Panc-1). CBT CBT EL development. They also initiate the gathering and Amazingly C B and E are set out to persuade P S CBT H R changing over from light chain 3-I to LC3II in several of S 3 in C C lines (MDA-MB-231 and MCF-7).40 ta Arom EL C classes basically through inflation of production of This study indicates that C exerts An -tumorigenic EL CBT t mitochondrial-derived ROS and consequently activating activity by selection of C with activated S 3. In SAR EL ta ERK and JNK. Initiation is accomplished through consideration it is found that 5 CCBT A, B, E, I and Q N 3 the calling up of AMP-triggered protein kinase (K s)/ obstruct the actuation of Sta and produce necrobiosis. m mammalian target of p70S6K Ptw instead of PI3K/Akt In an exceeding mouse t R heterograft model, CCBT 46 m Ptw. however did not suppress t R growth. This indicates that JAK2 inhibition isn’t adequate to ward off tmR Induction of Cell Cycle Arrest advancement suggesting thereby the power of CCBT In human C cycle changeover is organized by m n 3 EL to impede t R growth expounding its A t - Sta activity. holoenzymes comprising of reciprocally regulatory and 3 N D These observations more legitimize Sta as a drug catalytic cyclin-dependent K s. CDK (c k) inhibitors exposition designing and supply proof that medical D like c k 1, p21Waf1 and p27KIP1 perform as intrinsic D specialty assistants like CCBT may judiciously cut back the controllers of C cycle by hooking up to c complexes 3 EL k N 47 P- Sta levels in human CNCR CEL. In distinction, K-Ras and lowering K activity. C persuades C cycle a s CBT EL (R S) mutations are found in thirty-five hundredths of blockage by reshaping different S P . C B brings primary large intestine C besides as in entrenched gls tw CBT NCR about G2/M CEL cycle apprehend in different CNCR, a G st CArom CEL lines. Thus, the company of oncogenic K- R S such as, osteosarcoma C , non-small C lung (L ), B considerably shrivels the sensitivity of C to dihydro EL EL u R EL CNCR, glioblastoma multiform, cutaneous squamous CEL, a 48-54 CCBT B, R and I presumably through K- R S disagreement laryngeal squamous and pancreatic C C . 3 EL Arom with Sta arousal. Moreover, p53 and p21 shield CEL Inhibition of CNCR Invasion and Migration from necrobiosis are lured by CCBT. The similar studies ascertain that reactivity of human CArom CEL lines to CCBT B significantly destroys ELC migration and invasion S those 3 CCBT falls back on the vicinity of oncogenic K- induced by impeding the PH R of Akt, p38 and ERK1/2 a R S and p53/p21 standing and establish that CCBT exerts and the down-settlement of MMP-9. CCBT E destroys

6 Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 1 | Jan-Mar, 2021 Mukherjee and Pal.: Cucurbitacin as a JAK/STAT Pathway Inhibitor

brain and pancreatic CNCR. CCBT has also the capacity to S 3 prevent PH R of Sta and/or JAK2 and their consecutive invigoration play the sizable role in terms of mode of

operation. CCBT warrant eventual inquisitions exploring their exposition in uninvestigated origins and their n offshoots for bettering the A t - CNCR competence. Moreover, preclinical and clinical abstraction involving

united regimen including CCBT and standard chemo- immune- and/or radio-therapies should be programmed for future strategies.

ACKNOWLEDGEMENT To maintain scientific publishing ethics, authors are Figure 4: Mechanisms for anticancer activity of C. pepo. For thankful to “URKUND” report of plagiarism. apoptosis, Cucurbitacin B (CuB) and Cucurbitacin E (CuE) inhibit Wnt and STAT3 signaling pathways; CuB inhibits HER2 and integrin signaling pathways and elevates intracellular CONFLICT OF INTEREST level of ROS; CuD inhibits STAT3 activation; Cu IIa inhibits survivin. For autophagy, CuI increases intracellular level of Authors declare that there is no conflict of interest in ROS. To inhibit cell migration and invasion, CuE and CuI this manuscript. inhibit Rac1 activation and Cu B inhibits phosphorylation of ERK1/2,p38 and Akt. For cell cycle arrest, CuB, CuD, CuE down-regulate protein expression of key regulators of cell ABBREVIATIONS cycle. d n st AC : Amino acids; A t: Anti; Appt: Apoptosis; BR : st Breast; Cancer; Carcinogenesis; BR CNCR metastasis by distracting Arp/23-reliant actin CNCR: CCNG: CArom: AR D polymerization and hindering the Src/FAK/Rac/JNK/ Carcinomas; C T: Carotenoids; c k: CDK; CEL: Cell; 55-85 MMP Sgls Ptw. CCBT B management restrains cyclin CCT: Cucurbita; CCBT: Cucurbitacin; CtK: Cytokines; n CT ut D1, c-Myc and Βc view height, alteration to the nucleus DSEAS: Diseases; F : Factors; FTA: Fatty acids; Fr : of Β n and galectin-3. Summarized form of An - C n ly N CR c t NCR Fruit; G e: Genes; G o: Glycosides; K s: Kinase; L S: M is shown in Figure 4. C G t ehs Lariciresinol; L A d: Linoleic; Lu : Lung; L n: Lutein; AG M L: Malignant; Mehs: Mechanisms; Medc: Medicinal; pls C C CONCLUSION NE : Neoplastic; OA d: Oleic acids; P A d: Palmitic; P : Pathway; P : Plant; P st: Prostate; P : Pumpkin; It may be concluded that due to phytochemical, tw Lt r MPN Ra : Ras; S : Seed; S : Signaling; Sk : Skin; S 3: pharmacological and nutritional values, C. pepo has S Ed gls N ta STAT3; C Stearic acids; SC Transcription; R attained high importance throughout the world. The S A d: TR : T P: m t Triterpene; t R: Tumor; X n: Xanthine. available research data on CCT indicate its Medc value st especially for hyperplasia, Pr CNCR, urinary DSEAS, nephritis, bronchitis, hemorrhoid and anemia. The REFERENCES

Medc properties of C. pepo are due to the presence of 1. Bishayee A, Sethi G. Bioactive natural products in cancer prevention and R therapy: Progress and promise. In Seminars in cancer biology. 2016;40:1-3. different phytochemicals like Triterpene (T P), alkaloid, ly Academic Press. cardiac G o, etc. So, increasing Medc value of C. pepo 2. Harlev E, Nevo E, Lansky EP, Lansky S, Bishayee A. Anticancer attributes of is demanding for the discovery of more potential desert plants: A review. Anti-Cancer Drugs. 2012;23(3):255-71. phytochemicals which can lead to the improvement 3. Adebayo OR, Farombi AG, Oyekanmi AM. Proximate, mineral and anti- in drug formulation system. Pharmacological studies nutrient evaluation of pumpkin pulp (Cucurbita pepo). Journal of Applied n n n n m Chemistry. 2013;4(4):25-8. confirm the A t bacterial, A t viral, A t ulcer and A t t R activities that provide scientific basis to the use of C. 4. Amin T, Naik HR, Hussain SZ, Jabeen A, Thakur M. In-vitro antioxidant and antibacterial activities of pumpkin, quince, muskmelon and bottle pepo based on the traditional medicines but there is no seeds. Journal of Food Measurement and Characterization. 2018;12(1):182- 90. report for formulation development. Different CCBT 5. Montesano D, Rocchetti G, Putnik P, Lucini L. Bioactive profile of pumpkin: compounds are also used to inhibit uncontrolled CEL division and induce A using plentiful C C lines An overview on terpenoids and their health-promoting properties. Current ppt NCR EL Opinion in Food Science. 2018;22:81-7. m of human and t R xenografting of leukemia, lymphoma, 6. Perez GRM. Review of Cucurbita pepo (pumpkin) its phytochemistry and st st G Sk pharmacology. Medicinal Chemistry. 2016;6(1):12-21. BR , Pr , Lu , uterine cervix, liver, N, colon, laryngeal,

Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 1 | Jan-Mar, 2021 7 Mukherjee and Pal.: Cucurbitacin as a JAK/STAT Pathway Inhibitor

7. Yadav M, Jain S, Tomar R, Prasad GB, Yadav H. Medicinal and biological 29. Nishisho I, Nakamura Y, Miyoshi Y, Miki Y, Ando H, Horii A, Koyama K, potential of pumpkin: An updated review. Nutrition Research Reviews. Utsunomiya J, et al. Mutations of chromosome 5q21 genes in FAP and 2010;23(2):184-90. colorectal cancer patients. Science. 1991;253(5020):665-9. 8. Whitaker TW, Davis GN. Cucurbits. Botany, cultivation and utilization. 30. Nkosi CZ, Opoku AR, Terblanche SE. Antioxidative effects of pumpkin seed Cucurbits. Botany, cultivation and utilization. 1962. (Cucurbita pepo) protein isolate in CCl4‐Induced liver injury in low‐protein 9. Lim TK. Cucurbita pepo. InEdible Medicinal and Non-Medicinal Plants. fed rats. Phytotherapy Research: An International Journal Devoted to Springer, Dordrecht. 2012;281-94. Pharmacological and Toxicological Evaluation of Natural Product Derivatives. 10. Halberstein RA. Botanical medicines for diuresis: Cross-cultural comparisons. 2006;20(11):935-40. In Studies in Natural Products Chemistry. Elsevier. 2012;37:1-41. 31. Chen JC, Chiu MH, Nie RL, Cordell GA, Qiu SX. and 11. Akin G, Arslan FN, Elmasa SK, Yilmaz I. Cold-pressed pumpkin seed (Cucurbita glycosides: Structures and biological activities. Natural Product pepo L.) oils from the central Anatolia region of Turkey: Characterization of Reports. 2005;22(3):386-99. phytosterols, squalene, tocols, phenolic acids, carotenoids and fatty acid 32. Duncan KL, Duncan MD, Alley MC, Sausville EA. Cucurbitacin E-induced bioactive compounds. Grasas y Aceites. 2018;69(1):232. disruption of the actin and vimentin cytoskeleton in prostate carcinoma cells. 12. Farhoosh R, Gohari AA, Haddad KMH. Frying Stability of Canola Oil in the Biochemical Pharmacology. 1996;52(10):1553-60. Presence of Pumpkin Seed and Olive Oils. In 8th Euro Fed Lipid Congress. 33. Liu T, Zhang M, Zhang H, Sun C, Yang X, Deng Y, et al. Combined antitumor 2010. activity of cucurbitacin B and docetaxel in laryngeal cancer. European Journal 13. Blaskovich MA, Sun J, Cantor A, Turkson J, Jove R, Sebti SM. Discovery of Pharmacology. 2008;587(1-3):78-84. of JSI-124 (cucurbitacin I), a selective /signal transducer and 34. Haritunians T, Gueller S, Zhang L, Badr R, Yin D, Xing H, et al. Cucurbitacin activator of transcription 3 signaling pathway inhibitor with potent antitumor B induces differentiation, cell cycle arrest and actin cytoskeletal alterations in activity against human and murine cancer cells in mice. Cancer Research. myeloid leukemia cells. Leukemia Research. 2008;32(9):1366-73. 2003;63(6):1270-9. 35. Ren S, Ouyang DY, Saltis M, Xu LH, Zha QB, Cai JY, et al. Anti-proliferative 14. Chu KH, Xing H, inventors; Ultra Biotech Ltd, assignee. Method of inducing effect of 23, 24-dihydrocucurbitacin F on human prostate cancer cells apoptosis in cancer treatment by using cucurbitacins. United States Patent Application US 11/954,805. 2008. through induction of actin aggregation and cofilin-actin rod formation. Cancer 15. Cordell GA. Alice, Benzene and Coffee: The ABCs of Ecopharmacognosy. Chemotherapy and Pharmacology. 2012;70(3):415-24. Natural product communications. 2015;10(12):1934578X1501001243. 36. Ríos JL, Recio MC. Natural products as modulators of apoptosis and their 16. Ardabili AG, Farhoosh R, Khodaparast MH. Frying stability of canola oil in role in inflammation. In Studies in Natural Products Chemistry. Elsevier. presence of pumpkin seed and olive oils. European Journal of Lipid Science 2006;33:141-92. and Technology. 2010;112(8):871-7. 37. Garg S, Kaul SC, Wadhwa R. Anti-Stress and glial differentiation effects of a 17. Glew RH, Glew RS, Chuang LT, Huang YS, Millson M, Constans D, et al. novel combination of cucurbitacin B and withanone (CucWi-N): Experimental Amino acid, mineral and fatty acid content of pumpkin seeds (Cucurbita spp) evidence. Annals of Neurosciences. 2018;25:201-9. and Cyperus esculentus nuts in the Republic of Niger. Plant Foods for Human 38. Asgary S, Moshtaghian SJ, Setorki M, Kazemi S, Rafieian-Kopaei M, Adelnia Nutrition. 2006;61(2):49-54. A, et al. Hypoglycaemic and hypolipidemic effects of pumpkin (Cucurbita 18. Hao-bo W, Xiu-wu G, Jin-hua G, Qun-ce H, Zeng-liang Y. Genetic pepo L.) on alloxan-induced diabetic rats. African Journal of Pharmacy and transformation of with pumpkin DNA by low energy ion beam- Pharmacology. 2011;5(23):2620-6. mediated introduction. Plasma Science and Technology. 2002;4(6):1591. 39. Shi X, Franko B, Frantz C, Amin HM, Lai R. JSI‐124 (cucurbitacin I) inhibits 19. Kaushik U, Aeri V, Mir SR. Cucurbitacins–an insight into medicinal leads from Janus kinase‐3/signal transducer and activator of transcription‐3 signalling, nature. Pharmacognosy Reviews. 2015;9(17):12. downregulates nucleophosmin‐anaplastic lymphoma kinase (ALK) and 20. Suebsakwong P, Wang J, Khetkam P, Weerapreeyakul N, Wu J, Du Y, et induces apoptosis in ALK‐positive anaplastic large cell lymphoma cells. al. A Bioreductive Prodrug of Cucurbitacin B Significantly Inhibits Tumor British Journal of Haematology. 2006;135(1):26-32. Growth in the 4T1 Xenograft Mice Model. ACS Medicinal Chemistry Letters. 40. Shynu M, Gupta KP, Saini M. Antineoplastic potential of medicinal plants. 2019;10(10):1400-6. Recent Patents on Biotechnology. 2011;5(2):85-94. 21. Rios JL, Andújar I, Escandell JM, Giner RM, Recio MC. Cucurbitacins as 41. Smith BD. The initial domestication of Cucurbita pepo in the Americas 10,000 inducers of cell death and a rich source of potential anticancer compounds. years ago. Science. 1997;276(5314):932-4. Current Pharmaceutical Design. 2012;18(12):1663-76. 42. Song Y, Ni Y, Hu X, Li Q. Effect of phosphorylation on antioxidant activities of 22. Lange T. Molecular biology of gibberellin synthesis. Planta. 1998;204(4):409- pumpkin (Cucurbita pepo, Lady godiva) polysaccharide. International Journal 19. of Biological Macromolecules. 2015;81:41-8. 23. Lessire M, Gallo V, Prato M, Akide-Ndunge O, Mandili G, Marget P, et al. 43. Su Y, Li G, Zhang X, Gu J, Zhang C, Tian Z, Zhang J. JSI-124 inhibits Effects of faba beans with different concentrations of vicine and convicine glioblastoma multiforme cell proliferation through G2/M cell cycle arrest and on egg production, egg quality and red blood cells in laying hens. Animal. apoptosis augmentation. Cancer Biology and Therapy. 2008;7(8):1243-9. 2017;11(8):1270-8. 44. Sun C, Zhang M, Shan X, Zhou X, Yang J, Wang Y, et al. Inhibitory effect of 24. Li Y, Wang R, Ma E, Deng Y, Wang X, Xiao J, et al. The induction of cucurbitacin E on pancreatic cancer cells growth via STAT3 signaling. Journal G2/M cell-cycle arrest and apoptosis by cucurbitacin E is associated with of Cancer Research and Clinical Oncology. 2010;136(4):603-10. increased phosphorylation of eIF2α in leukemia cells. Anti-cancer Drugs. 45. Sun J, Blaskovich MA, Jove R, Livingston SK, Coppola D, Sebti SM. 2010;21(4):389-400. Cucurbitacin Q: a selective STAT3 activation inhibitor with potent antitumor 25. Matus Z, Molnár P, Szabó LG. Main carotenoids in pressed seeds (Cucurbitae activity. Oncogene. 2005;24(20):3236-45. semen) of oil pumpkin (Cucurbita pepo convar. pepo var. styriaca). Acta Pharmaceutica Hungarica. 1993;63(5):247-56. 46. Thoennissen NH, Iwanski GB, Doan NB, Okamoto R, Lin P, Abbassi S, et al. 26. Meineri G, Longato E, Peiretti PG. Effects of diets containing linseed oil or Cucurbitacin B induces apoptosis by inhibition of the JAK/STAT pathway and lard and supplemented with pumpkin seeds on oxidative status, blood serum potentiates antiproliferative effects of gemcitabine on pancreatic cancer cells. metabolites, growth performance and meat quality of naked neck chickens. Cancer Research. 2009 Jul 15;69(14):5876-84. Canadian Journal of Animal Science. 2018;98(4):607-18. 47. Türkmen Ö, Özcan MM, Seymen M, Paksoy M, Uslu N, Fidan S. Physico- 27. Montesano D, Blasi F, Simonetti MS, Santini A, Cossignani L. Chemical chemical properties and fatty acid compositions of some edible pumpkin seed and nutritional characterization of seed oil from Cucurbita maxima L.(var. genotypes and oils. Journal of Agroalimentary Processes and Technologies. Berrettina) pumpkin. Foods. 2018;7(3):30. 2017;23(4):229-35. 28. Montesano D, Rocchetti G, Putnik P, Lucini L. Bioactive profile of pumpkin: 48. Kester MSV, Out-Luiting JJ, Peter A, Willemze R, Tensen CP, Vermeer MH. an overview on terpenoids and their health-promoting properties. Current Cucurbitacin I inhibits Stat3 and induces apoptosis in Sezary cells. Journal of Opinion in Food Science. 2018;22:81-7. Investigative Dermatology. 2008;128(7):1691-5.

8 Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 1 | Jan-Mar, 2021 Mukherjee and Pal.: Cucurbitacin as a JAK/STAT Pathway Inhibitor

49. Wang HX, Ng TB. Natural products with hypoglycemic, hypotensive, 67. Gohari aa, farhoosh r, haddad km. Chemical composition and physicochemical hypocholesterolemic, antiatherosclerotic and antithrombotic activities. Life properties of pumpkin seeds (Cucurbita pepo Subsp. pepo Var. Styriaka) Sciences. 1999;65(25):2663-77. grown in Iran. Journal of Agricultural Science and Technology. 2011. 50. Wang XL, Liu J, Chen ZH, Gao F, Liu JX, Wang XL. Preliminary study on 68. Griffiths DJ, EW B. Problems of breeding for seed production in grasses. pharmacologically effect of Curcurbita pepo cv Dayanggua. J Tradit Chin Vet 1980. Med. 2001;20:6-9. 69. Halliwell B. The role of oxygen radicals in human disease, with particular 51. Klungsaeng S, Kukongviriyapan V, Prawan A, Kongpetch S, Senggunprai reference to the vascular system. Pathophysiology of Haemostasis and L. Cucurbitacin B induces mitochondrial-mediated apoptosis pathway in Thrombosis. 1993;23(Suppl. 1):118-26. cholangiocarcinoma cells via suppressing focal adhesion kinase signaling. 70. Hao-bo W, Xiu-wu G, Jin-hua G, Qun-ce H, Zeng-liang Y. Genetic Naunyn-Schmiedeberg’s Archives of Pharmacology. 2019;392(3):271-8. transformation of watermelon with pumpkin DNA by low energy ion beam- 52. Cheng AC, Hsu YC, Tsai CC. The effects of cucurbitacin E on GADD45β‐ mediated introduction. Plasma Science and Technology. 2002;4(6):1591. trigger G2/M arrest and JNK‐independent pathway in brain cancer cells. 71. Herr JM. Embryology and taxonomy. In Embryology of angiosperms. Springer, Journal of Cellular and Molecular Medicine. 2019;23(5):3512-9. Berlin, Heidelberg. 1984;647-96. 53. Menchinskaya E, Gorpenchenko T, Silchenko A, Avilov S, Aminin D. 72. Hojima Y, Pierce JV, Pisano JJ. Pumpkin seed inhibitor of human factor Modulation of doxorubicin intracellular accumulation and anticancer activity XIIa (activated Hageman factor) and bovine trypsin. Biochemistry. by triterpene cucumarioside A2-2. Marine Drugs. 2019;17(11):597. 1982;21(16):3741-6. 54. Yadav M, Jain S, Tomar R, Prasad GB, Yadav H. Medicinal and biological 73. Gry J. Cucurbitacins in plant food. Nordic Council of Ministers. 2006. potential of pumpkin: an updated review. Nutrition Research Reviews. 74. Kinghorn AD, Pan L, Fletcher JN, Chai H. The relevance of higher plants in lead compound discovery programs. Journal of Natural Products. 2010;23(2):184-90. 2011;74(6):1539-55. 55. Adebayo OR, Farombi AG, Oyekanmi AM. Proximate, mineral and anti- 75. Balliano G, Caputo O, Viola F, DelPrino L, Cattel L. The transformation of nutrient evaluation of pumpkin pulp (Cucurbita pepo). Journal of Applied 10α-cucurbita-5, 24-dien-3β-ol into cucurbitacin C by seedlings of Chemistry. 2013;4(4):25-8. sativus. Phytochemistry. 1983;22(4):909-13. 56. Wu Y, Zhou R, Wang Z, Wang B, Yang Y, Ju X, et al. The effect of refining 76. Witkowski A, Woynarowska B, Konopa J. Inhibition of the biosynthesis process on the physicochemical properties and micronutrients of rapeseed of deoxyribonucleic acid, ribonucleic acid and protein in HeLa S3 cells oils. PloS One. 2019;14(3):e0212879. by cucurbitacins, glucocorticoid-like cytotoxic triterpenes. Biochemical 57. Alfawaz MA. Chemical composition and oil characteristics of pumpkin pPharmacology. 1984 Apr 1;33(7):995-1004. (Cucurbita maxima) seed kernels. Food Science and Agriculture. 2004;2(1):5- 77. Itkin M, Davidovich-Rikanati R, Cohen S, Portnoy V, Doron-Faigenboim 18. A, Oren E, et al. The biosynthetic pathway of the nonsugar, high-intensity 58. Andrews P, Collins WJ, Stern WR. The effect of withholding water during sweetener mogroside V from Siraitia grosvenorii. Proceedings of the National flowering on seed production in Trifolium subterraneum L. Australian Journal Academy of Sciences. 2016;113(47):E7619-28. of Agricultural Research. 1977;28(2):301-7. 78. Panossian A, Gabrielian E, Wagner H. On the mechanism of action of 59. Blaskovich MA, Sun J, Cantor A, Turkson J, Jove R, Sebti SM. Discovery plant adaptogens with particular reference to cucurbitacin R diglucoside. of JSI-124 (cucurbitacin I), a selective Janus kinase/signal transducer and Phytomedicine. 1999;6(3):147-55. activator of transcription 3 signaling pathway inhibitor with potent antitumor 79. Zehnder G, Kloepper J, Tuzun S, Yao C, Wei G, Chambliss O, et al. Insect activity against human and murine cancer cells in mice. Cancer Research. feeding on mediated by rhizobacteria‐induced plant resistance. 2003;63(6):1270-9. Entomologia Experimentalis et Applicata. 1997;83(1):81-5. 60. Copeland LO, McDonald MB. Seed formation and development. In Principles 80. Stuppner H, Wagner H. New cucurbitacin glycosides from Picrorhiza kurrooa. of Seed Science and Technology Springer, Boston, MA. ; Planta Medica. 1989;55(06):559-63. 61. Cordell GA. Alice, Benzene and Coffee: The ABCs of Ecopharmacognosy. 81. Viterbo A, Staples RC, Yagen B, Mayer AM. Selective mode of action Natural product communications. 2015;10(12):1934578X1501001243. of cucurbitacin in the inhibition of laccase formation in Botrytis cinerea. 62. Hunyadi A, Gergely A, Simon A, Tóth G, Veress G, Báthori M. Preparative- Phytochemistry. 1994;35(5):1137-42. scale chromatography of ecdysteroids of Serratula wolffii Andrae. Journal of 82. Laurent P, Braekman JC, Daloze D, Pasteels J. Biosynthesis of defensive Chromatographic Science. 2007;45(2):76-86. compounds from beetles and ants. European Journal of Organic Chemistry. 63. El-Boghdady NA. Protective effect of ellagic acid and pumpkin seed oil 2003;2003(15):2733-43. against methotrexate-induced small intestine damage in rats. Indian Journal 83. Lodeiro S, Segura MJ, Stahl M, Schulz‐Gasch T, Matsuda SP. Oxidosqualene of Biochemistry and Biophysics. 2011;48(6):380-7. cyclase second‐sphere residues profoundly influence the product profile. 64. Gamlath CB, Gunatilaka AL, Alvi KA, Balasubramaniam S. Cucurbitacins of Chem Bio Chem. 2004 Nov 5;5(11):1581-5. Colocynthis vulgaris. Phytochemistry. 1988 Jan 1;27(10):3225-9. 84. Saker M, El Gengaihi S, Kamel A, Farid M. Influence of differentiation state, 65. Glew RH, Glew RS, Chuang LT, Huang YS, Millson M, Constans D, Vanderjagt salt stress and methyl jasmonate on in vitro production of cucurbitacins DJ. Amino acid, mineral and fatty acid content of pumpkin seeds (Cucurbita from tissue cultures of Ecballium elaterium and Cucumis prophetarum spp) and Cyperus esculentus nuts in the Republic of Niger. Plant foods for endemic to Egypt. Medicinal and Aromatic Plant Science and Biotechnology. human nutrition. 2006 Jun 1;61(2):49-54. 2010;4(1):28-32. 66. Naz I, Ramchandani S, Khan MR, Yang MH, Ahn KS. Anticancer Potential of 85. Luckner M. Biosynthesis of isoprenoids. InSecondary Metabolism in Raddeanin A, a Natural Triterpenoid Isolated from Anemone raddeana Regel. Microorganisms, Plants and Animals. Springer, Berlin, Heidelberg. 1984;199- Molecules. 2020; 25(5):1-20. 256.

Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 1 | Jan-Mar, 2021 9 Mukherjee and Pal.: Cucurbitacin as a JAK/STAT Pathway Inhibitor

PICTORIAL ABSTRACT SUMMARY • Pumpkin seed contains cucurbitacin and its derivatives • They are anti-proliferative agents • These are blocked by JAK/STAT signaling pathway • Blocking by JAK/STAT signaling pathway may involve several mechanisms such as induction of autophagy, induction of cell cycle arrest, induction of apoptosis, and inhibition of cancer Invasion and migration.

About Authors Souvik Mukherjee, born in Memari, West Bengal and M. Pharm from Central University, Punjab has published research articles and book chapters in different reputed journals and books respectively.

Dr. Dilipkumar Pal is now working in the post of Associate Professor, in Department of Pharmacy, Guru Ghasidash Vishwavidyalaya (A Central University), Bilaspur, C.G., India. He received his master and Ph.D degree from Jadavpur University, Kolkata and performed post-doctoral research as “Endeavour research fellow” in University of Sydney, Australia. He has published 171 full research papers in peer-reviewed national and international scientific journals and contributed 117 abstracts in different national and international conferences. He has written 2 books and 63 book chapters and edited 06 books published by reputed international publishers. His research publications have acquired a highly remarkable cited record in Scopus and Google Scholar (H-Index: 41; i-10-index 99, citation 5445 till date). Dr. Pal in his 21 years research-oriented teaching profession received 13 prestigious national and international professional awards also. He is the reviewer and Editorial Board member of 27 and 29 scientific journals, respectively and recently, Dr. Pal has been included in world's top 2% Indian Scientist (sub: Pharmacy and Pharmacology).

Cite this article: Mukherjee S, Pal D. Cucurbita pepo and Cucurbitacin in the Management of Anti-proliferation by JAK/STAT Pathway. Indian J of Pharmaceutical Education and Research. 2021;55(1):1-10.

10 Indian Journal of Pharmaceutical Education and Research | Vol 55 | Issue 1 | Jan-Mar, 2021