Phorbol is a natural product found in many plants, especially those of the Euphorbiaceae and Thymelaeaceae families.[10][11] Phorbol is the active constituent of the highly toxic New World tropical manchineel or beach apple, Hippomane mancinella.[12] It is very soluble in most polar organic solvents, as well as in water. In the manchineel, this leads to an additional exposure risk during rain, where liquid splashing from an undamaged tree may also be injurious. Contact with the tree or consumption of its fruit can lead to symptoms such as severe pain and swelling.[13][14]
The purging croton, Croton tiglium, is the source of croton oil from which phorbol was initially isolated. Its seeds and oil have been used for hundreds of years in traditional medicine, generally as a purgative, and the seeds were mentioned in Chinese herbal texts 2000 years ago.[15] The purgative effects of the oil are largely attributed to the high percentage of phorbol esters contained in the oil. Phorbol was isolated from C. tiglium seeds in 1934.[2][3][4][5][6] The structure of the compound was determined in 1967,[7][8] and a total synthesis was described in 2015.[16]
Mechanism of action
Phorbol derivatives work primarily by interacting with protein kinase C (PKC), although they can interact with other phospholipid membrane receptors.[17] The esters bind to PKC in a similar way to its natural ligand, diacylglycerol, and activate the kinase.[18] Diacylglycerol is degraded quickly by the body, allowing PKC to be reversibly activated. When phorbol esters bind to the receptor, they are not degraded as efficiently by the body, leading to constitutively active PK.[17] PKC is involved in a number of important cell signaling pathways. Thus, phorbol ester exposure can show a wide range of results.
Crystal structure of phorbol-13-acetate bound to the C1B domain of protein kinase C delta
The main results of phorbol exposure are tumor promotion and inflammatory response. Although phorbol is not a carcinogen itself, it greatly enhances the action of other substances and promotes tumor proliferation. PKC is a key component in biological pathways controlling cell growth and differentiation. When phorbol esters bind to PKC, cell proliferation pathways are activated. This effect greatly promotes tumors when the cells are exposed to even a sub-carcinogenic amount of a substance.[17] PKC is also involved in activation of inflammation pathways such as the NF-κB pathway. Thus, exposure to phorbol products can induce an inflammatory response in tissues.[19][20] Symptoms can include edema and pain, especially to the skin and mucus membranes.[10] While phorbol itself does not have irritant activity, nearly all phorbol esters are highly irritant, with a wide range of half-maximal inhibitory concentration (IC50) values.[10] The median lethal dose (LD50) of phorbol esters for male mice was found to be about 27 mg/kg, with the mice showing hemorrhage and congestion of pulmonary blood vessels, as well as lesions throughout the body.[18]
Total synthesis
A total synthesis of enantiopure phorbol was developed in 2015. While this synthesis will not replace natural isolation products, it will enable researchers to create phorbol analogs for use in research, especially creating phorbol derivatives that can be evaluated for anti-cancer activity.[16] Previously, the difficulty with synthesizing phorbol had been creating C–C bonds, especially in the six-membered ring at the top of the molecule. This synthesis starts from (+)-3-carene, and uses a series of 19 steps to eventually create (+)-phorbol.[21][22][16]
Overview of the complete synthesis of (+)-phorbol starting with (+)-3-carene
Uses in biomedical research and treatments
Because of their mechanism of action, phorbol esters can be used to study tumor proliferation and pain response. TPA is most commonly used in the laboratory to induce a cellular response. For example, TPA can be used to measure response to pain and test compounds that may mitigate the inflammatory response.[23] TPA and other phorbol esters can also be used to induce tumor formation and elucidate its mechanism.[10] Some phorbol esters such as tigilanol tiglate have shown anti-cancer activities and can potentially be used as a defense against certain viruses and bacteria.[10] TPA, together with ionomycin, can also be used to stimulate T-cell activation, proliferation, and cytokine production, and is used in protocols for intracellular staining of these cytokines.[24] Phorbol, in the form of croton oil, is also in folk medicine as a purgative, counter-irritant, or anthelmintic.[25][26]
^ abFlaschenträger B; v. Wolffersdorff R (1934). "Über den Giftstoff des Crotonöles. 1. Die Säuren des Crotonöles". Helvetica Chimica Acta. 17 (1): 1444–1452. doi:10.1002/hlca.193401701179.
^ abFlaschenträger B, Wigner G (1942). "Über den Giftstoff des Crotonöles. V. Die Gewinnung von Crotonharz, Dünnem Öl und Phorbol aus dem Crotonöl durch Alkoholyse". Helvetica Chimica Acta. 25 (3): 569–581. doi:10.1002/hlca.19420250315.
^ abKauffmann T, Neumann H, Lenhardt K (1959). "Zur Konstitution des Phorbols, I. Über die reduzierende Gruppe des Phorbols". Chemische Berichte. 92 (8): 1715–1726. doi:10.1002/cber.19590920802.
^ abKauffmann T, Eisinger A, Jasching W, Lenhardt K (1959). "Zur Konstitution des Phorbols, I. Über die reduzierende Gruppe des Phorbols". Chemische Berichte. 92 (8): 1727–1738. doi:10.1002/cber.19590920803.
^ abTseng SS, van Duuren BL, Solomon JJ (1977). "Synthesis of 4aα-Phorbol 9-Myristate 9a-Acetate and Related Esters". J. Org. Chem. 42 (33): 3645–3649. doi:10.1021/jo00443a002. PMID 915585.
^ abHecker E; Bartsch H; Bresch H; Gschwendt M; Härle B; Kreibich G; Kubinyi H; Schairer HU; v. Szczepanski C; Thielmann HW (1967). "Structure and Stereochemistry of the Tetracyclic Diterpene Phorbol from Croton tiglium L". Tetrahedron Letters. 8 (33): 3165–3170. doi:10.1016/S0040-4039(01)89890-7.
^ abPettersen RC, Ferguson G, Crombie L, Games ML, Pointer DJ (1967). "The Structure and Stereochemistry of Phorbol, Diterpene Parent of Co-carcinogens of Croton Oil". Chem. Commun. 1967 (14): 716–717. doi:10.1039/C19670000716.
^Blumberg PM (1988). "Protein Kinase C as the Receptor for the Phorbol Ester Tumor Promoters: Sixth Rhoads Memorial Award Lecture" (PDF). Cancer Res. 48 (1): 1–8. PMID 3275491.
^ abcdeWang, Xiao-Yang; Liu, Li-Ping; Qin, Guo-Wei; Kang, Ting-Guo (2015). "Tigliane Diterpenoids from the Euphorbiaceae and Thymelaeaceae Families". Chemical Reviews. 115 (9): 2975–3011. doi:10.1021/cr200397n. PMID 25906056.
^Beutler, John A.; Alvarado, Ada Belinda; McCloud, Thomas G. (1989). "Distribution of phorbol ester bioactivity in the euphorbiaceae". Phytotherapy Research. 3 (5): 188–192. doi:10.1002/ptr.2650030507. S2CID 85408071.
^Adolf, W; Hecker, E (1984). "On the active principles of the spurge family. X. Skin irritants, cocarcingoens, and cryptic carcinogens from the latex of the manchineel tree". J Nat Prod. 47 (3): 482–496. doi:10.1021/np50033a015. PMID 6481361.
^Strickland, Nicola H (August 12, 2000). "Eating a manchineel "beach apple"". BMJ. 321 (7258): 428. doi:10.1136/bmj.321.7258.428. PMC1127797. PMID 10938053.
^Blue, Lauren M; Sailing, Christopher; DeNapoles, Christopher; Fondots, Jordan; Johnson, Edward S (2011). "Manchineel Dermatitis in North American Students in the Caribbean". Journal of Travel Medicine. 18 (6): 422–424. doi:10.1111/j.1708-8305.2011.00568.x. PMID 22017721.
^ abcKawamura, Shuhei; Chu, Hang; Felding, Jakob; Baran, Phil S. (2016). "Nineteen-step total synthesis of (+)-phorbol". Nature. 532 (7597): 90–3. Bibcode:2016Natur.532...90K. doi:10.1038/nature17153. PMC4833603. PMID 27007853..
^ abcGoel, Gunjan; Makkar, Harinder P.S.; Francis, George; Becker, Klaus (2007). "Phorbol Esters: Structure, Biological Activity, and Toxicity in Animals". International Journal of Toxicology. 26 (4): 279–288. CiteSeerX10.1.1.320.6537. doi:10.1080/10915810701464641. PMID 17661218. S2CID 11550625.
^ abLi, Cai-Yan; Devappa, Rakshit K; Liu, Jian-Xin; Lv, Jian-Min; Makkar, HPS; Becker, K (February 2010). "Toxicity of Jatropha curcas phorbol esters in mice". Food and Chemical Toxicology. 48 (2): 620–625. doi:10.1016/j.fct.2009.11.042. PMID 19944127.
^Moscat, Jorge; Diaz-Meco, María T; Rennert, Paul (January 2003). "NF-κB activation by protein kinase C isoforms and B-cell function". EMBO Reports. 4 (1): 31–36. doi:10.1038/sj.embor.embor704. PMC1315804. PMID 12524517.
^"NF-κB and Inflammation". Sigma-Aldrich. Sigma-Aldrich. Retrieved 7 May 2017.
^Wender, Paul A.; Kogen, Hiroshi; Lee, Hee Yoon; Munger, John D.; Wilhelm, Robert S.; Williams, Peter D. (1989). "Studies on tumor promoters. 8. The synthesis of phorbol". Journal of the American Chemical Society. 111 (24): 8957. doi:10.1021/ja00206a050.
^Wender, Paul A.; Rice, Kenneth D.; Schnute, Mark E. (1997). "The First Formal Asymmetric Synthesis of Phorbol". Journal of the American Chemical Society. 119 (33): 7897. doi:10.1021/ja9706256.
^Medeiros, Rodrigo; Otuki, Michel F; Avellar, Maria Christina W; Calixto, João (March 2007). "Mechanisms underlying the inhibitory actions of the pentacyclic triterpene α-amyrin in the mouse skin inflammation induced by phorbol ester 12-O-tetradecanoylphorbol-13-acetate". European Journal of Pharmacology. 22 (2–3): 227–235. doi:10.1016/j.ejphar.2006.12.005.
^"Flow Cytometry Intracellular Staining Guide". eBioscience, Inc. Retrieved 2011-09-25.
^Pal, Prince Kumar; Nandi, Manmath Kumar; Singh, Narendra Kumar (Jan–Mar 2014). "Detoxification of Croton tiglium L. seeds by Ayurvedic process of Śodhana". Ancient Science of Life. 33 (3): 157–161. doi:10.4103/0257-7941.144619. PMC4264303. PMID 25538350.
phorbol, natural, plant, derived, organic, compound, member, tigliane, family, diterpenes, first, isolated, 1934, hydrolysis, product, croton, which, derived, from, seeds, purging, croton, croton, tiglium, structure, phorbol, determined, 1967, various, esters,. Phorbol is a natural plant derived organic compound It is a member of the tigliane family of diterpenes Phorbol was first isolated in 1934 as the hydrolysis product of croton oil which is derived from the seeds of the purging croton Croton tiglium 2 3 4 5 6 The structure of phorbol was determined in 1967 7 8 Various esters of phorbol have important biological properties the most notable of which is the capacity to act as tumor promoters through activation of protein kinase C 9 They mimic diacylglycerols glycerol derivatives in which two hydroxyl groups have reacted with fatty acids to form esters The most common and potent phorbol ester is 12 O tetradecanoylphorbol 13 acetate TPA also called phorbol 12 myristate 13 acetate PMA which is used as a biomedical research tool in contexts such as models of carcinogenesis Phorbol 1 NamesPreferred IUPAC name 1aR 1bS 4aR 7aS 9bS 8R 9R 9aS 4a 7b 9 9a Tetrahydroxy 3 hydroxymethyl 1 1 6 8 tetramethyl 1 1a 1b 4 4a 7a 7b 8 9 9a decahydro 5H cyclopropa 3 4 benzo 1 2 e azulen 5 oneIdentifiersCAS Number 17673 25 5 Y3D model JSmol Interactive imageChEBI CHEBI 8116 NChemSpider 390610 NECHA InfoCard 100 162 035PubChem CID 442070UNII XUZ76S9127 YCompTox Dashboard EPA DTXSID2021155InChI InChI 1S C20H28O6 c1 9 5 13 18 24 15 9 22 7 11 8 21 6 12 14 17 3 4 20 14 26 16 23 10 2 19 12 13 25 h5 6 10 12 14 16 21 23 26H 7 8H2 1 4H3 t10 12 13 14 16 18 19 20 m1 s1 NKey QGVLYPPODPLXMB UBTYZVCOSA N NInChI 1 C20H28O6 c1 9 5 13 18 24 15 9 22 7 11 8 21 6 12 14 17 3 4 20 14 26 16 23 10 2 19 12 13 25 h5 6 10 12 14 16 21 23 26H 7 8H2 1 4H3 t10 12 13 14 16 18 19 20 m1 s1Key QGVLYPPODPLXMB UBTYZVCOBRSMILES OCC1 C C C C C 4C H C 4 O C H O C H 2C H C 2 O C C C C C3 O H C 3 O C1PropertiesChemical formula C 20H 28O 6Molar mass 364 438 g mol 1Melting point 250 to 251 C 482 to 484 F 523 to 524 K Solubility in water Soluble in DMSO 25mg ml 100 ethanol 25mg ml acetone ether or dimethyl formamide almost insoluble in aqueous buffers Except where otherwise noted data are given for materials in their standard state at 25 C 77 F 100 kPa N verify what is Y N Infobox references Contents 1 History and source 2 Mechanism of action 3 Total synthesis 4 Uses in biomedical research and treatments 5 References 6 External linksHistory and source EditPhorbol is a natural product found in many plants especially those of the Euphorbiaceae and Thymelaeaceae families 10 11 Phorbol is the active constituent of the highly toxic New World tropical manchineel or beach apple Hippomane mancinella 12 It is very soluble in most polar organic solvents as well as in water In the manchineel this leads to an additional exposure risk during rain where liquid splashing from an undamaged tree may also be injurious Contact with the tree or consumption of its fruit can lead to symptoms such as severe pain and swelling 13 14 The purging croton Croton tiglium is the source of croton oil from which phorbol was initially isolated Its seeds and oil have been used for hundreds of years in traditional medicine generally as a purgative and the seeds were mentioned in Chinese herbal texts 2000 years ago 15 The purgative effects of the oil are largely attributed to the high percentage of phorbol esters contained in the oil Phorbol was isolated from C tiglium seeds in 1934 2 3 4 5 6 The structure of the compound was determined in 1967 7 8 and a total synthesis was described in 2015 16 Mechanism of action EditPhorbol derivatives work primarily by interacting with protein kinase C PKC although they can interact with other phospholipid membrane receptors 17 The esters bind to PKC in a similar way to its natural ligand diacylglycerol and activate the kinase 18 Diacylglycerol is degraded quickly by the body allowing PKC to be reversibly activated When phorbol esters bind to the receptor they are not degraded as efficiently by the body leading to constitutively active PK 17 PKC is involved in a number of important cell signaling pathways Thus phorbol ester exposure can show a wide range of results Crystal structure of phorbol 13 acetate bound to the C1B domain of protein kinase C delta The main results of phorbol exposure are tumor promotion and inflammatory response Although phorbol is not a carcinogen itself it greatly enhances the action of other substances and promotes tumor proliferation PKC is a key component in biological pathways controlling cell growth and differentiation When phorbol esters bind to PKC cell proliferation pathways are activated This effect greatly promotes tumors when the cells are exposed to even a sub carcinogenic amount of a substance 17 PKC is also involved in activation of inflammation pathways such as the NF kB pathway Thus exposure to phorbol products can induce an inflammatory response in tissues 19 20 Symptoms can include edema and pain especially to the skin and mucus membranes 10 While phorbol itself does not have irritant activity nearly all phorbol esters are highly irritant with a wide range of half maximal inhibitory concentration IC50 values 10 The median lethal dose LD50 of phorbol esters for male mice was found to be about 27 mg kg with the mice showing hemorrhage and congestion of pulmonary blood vessels as well as lesions throughout the body 18 Total synthesis EditA total synthesis of enantiopure phorbol was developed in 2015 While this synthesis will not replace natural isolation products it will enable researchers to create phorbol analogs for use in research especially creating phorbol derivatives that can be evaluated for anti cancer activity 16 Previously the difficulty with synthesizing phorbol had been creating C C bonds especially in the six membered ring at the top of the molecule This synthesis starts from 3 carene and uses a series of 19 steps to eventually create phorbol 21 22 16 Overview of the complete synthesis of phorbol starting with 3 careneUses in biomedical research and treatments EditBecause of their mechanism of action phorbol esters can be used to study tumor proliferation and pain response TPA is most commonly used in the laboratory to induce a cellular response For example TPA can be used to measure response to pain and test compounds that may mitigate the inflammatory response 23 TPA and other phorbol esters can also be used to induce tumor formation and elucidate its mechanism 10 Some phorbol esters such as tigilanol tiglate have shown anti cancer activities and can potentially be used as a defense against certain viruses and bacteria 10 TPA together with ionomycin can also be used to stimulate T cell activation proliferation and cytokine production and is used in protocols for intracellular staining of these cytokines 24 Phorbol in the form of croton oil is also in folk medicine as a purgative counter irritant or anthelmintic 25 26 References Edit Merck Index 11th Edition 7306 a b Flaschentrager B v Wolffersdorff R 1934 Uber den Giftstoff des Crotonoles 1 Die Sauren des Crotonoles Helvetica Chimica Acta 17 1 1444 1452 doi 10 1002 hlca 193401701179 a b Flaschentrager B Wigner G 1942 Uber den Giftstoff des Crotonoles V Die Gewinnung von Crotonharz Dunnem Ol und Phorbol aus dem Crotonol durch Alkoholyse Helvetica Chimica Acta 25 3 569 581 doi 10 1002 hlca 19420250315 a b Kauffmann T Neumann H Lenhardt K 1959 Zur Konstitution des Phorbols I Uber die reduzierende Gruppe des Phorbols Chemische Berichte 92 8 1715 1726 doi 10 1002 cber 19590920802 a b Kauffmann T Eisinger A Jasching W Lenhardt K 1959 Zur Konstitution des Phorbols I Uber die reduzierende Gruppe des Phorbols Chemische Berichte 92 8 1727 1738 doi 10 1002 cber 19590920803 a b Tseng SS van Duuren BL Solomon JJ 1977 Synthesis of 4aa Phorbol 9 Myristate 9a Acetate and Related Esters J Org Chem 42 33 3645 3649 doi 10 1021 jo00443a002 PMID 915585 a b Hecker E Bartsch H Bresch H Gschwendt M Harle B Kreibich G Kubinyi H Schairer HU v Szczepanski C Thielmann HW 1967 Structure and Stereochemistry of the Tetracyclic Diterpene Phorbol from Croton tiglium L Tetrahedron Letters 8 33 3165 3170 doi 10 1016 S0040 4039 01 89890 7 a b Pettersen RC Ferguson G Crombie L Games ML Pointer DJ 1967 The Structure and Stereochemistry of Phorbol Diterpene Parent of Co carcinogens of Croton Oil Chem Commun 1967 14 716 717 doi 10 1039 C19670000716 Blumberg PM 1988 Protein Kinase C as the Receptor for the Phorbol Ester Tumor Promoters Sixth Rhoads Memorial Award Lecture PDF Cancer Res 48 1 1 8 PMID 3275491 a b c d e Wang Xiao Yang Liu Li Ping Qin Guo Wei Kang Ting Guo 2015 Tigliane Diterpenoids from the Euphorbiaceae and Thymelaeaceae Families Chemical Reviews 115 9 2975 3011 doi 10 1021 cr200397n PMID 25906056 Beutler John A Alvarado Ada Belinda McCloud Thomas G 1989 Distribution of phorbol ester bioactivity in the euphorbiaceae Phytotherapy Research 3 5 188 192 doi 10 1002 ptr 2650030507 S2CID 85408071 Adolf W Hecker E 1984 On the active principles of the spurge family X Skin irritants cocarcingoens and cryptic carcinogens from the latex of the manchineel tree J Nat Prod 47 3 482 496 doi 10 1021 np50033a015 PMID 6481361 Strickland Nicola H August 12 2000 Eating a manchineel beach apple BMJ 321 7258 428 doi 10 1136 bmj 321 7258 428 PMC 1127797 PMID 10938053 Blue Lauren M Sailing Christopher DeNapoles Christopher Fondots Jordan Johnson Edward S 2011 Manchineel Dermatitis in North American Students in the Caribbean Journal of Travel Medicine 18 6 422 424 doi 10 1111 j 1708 8305 2011 00568 x PMID 22017721 Zhang Xiao Long Wang Lun Li Fu Yu Kai Wang Ming Kui 2013 Cytotoxic Phorbol Esters of Croton tiglium Journal of Natural Products 13 5 858 864 doi 10 1021 np300832n PMID 23701597 a b c Kawamura Shuhei Chu Hang Felding Jakob Baran Phil S 2016 Nineteen step total synthesis of phorbol Nature 532 7597 90 3 Bibcode 2016Natur 532 90K doi 10 1038 nature17153 PMC 4833603 PMID 27007853 a b c Goel Gunjan Makkar Harinder P S Francis George Becker Klaus 2007 Phorbol Esters Structure Biological Activity and Toxicity in Animals International Journal of Toxicology 26 4 279 288 CiteSeerX 10 1 1 320 6537 doi 10 1080 10915810701464641 PMID 17661218 S2CID 11550625 a b Li Cai Yan Devappa Rakshit K Liu Jian Xin Lv Jian Min Makkar HPS Becker K February 2010 Toxicity of Jatropha curcas phorbol esters in mice Food and Chemical Toxicology 48 2 620 625 doi 10 1016 j fct 2009 11 042 PMID 19944127 Moscat Jorge Diaz Meco Maria T Rennert Paul January 2003 NF kB activation by protein kinase C isoforms and B cell function EMBO Reports 4 1 31 36 doi 10 1038 sj embor embor704 PMC 1315804 PMID 12524517 NF kB and Inflammation Sigma Aldrich Sigma Aldrich Retrieved 7 May 2017 Wender Paul A Kogen Hiroshi Lee Hee Yoon Munger John D Wilhelm Robert S Williams Peter D 1989 Studies on tumor promoters 8 The synthesis of phorbol Journal of the American Chemical Society 111 24 8957 doi 10 1021 ja00206a050 Wender Paul A Rice Kenneth D Schnute Mark E 1997 The First Formal Asymmetric Synthesis of Phorbol Journal of the American Chemical Society 119 33 7897 doi 10 1021 ja9706256 Medeiros Rodrigo Otuki Michel F Avellar Maria Christina W Calixto Joao March 2007 Mechanisms underlying the inhibitory actions of the pentacyclic triterpene a amyrin in the mouse skin inflammation induced by phorbol ester 12 O tetradecanoylphorbol 13 acetate European Journal of Pharmacology 22 2 3 227 235 doi 10 1016 j ejphar 2006 12 005 Flow Cytometry Intracellular Staining Guide eBioscience Inc Retrieved 2011 09 25 Croton Oil OilHealthBenefits OilHealthBenefits 2014 07 12 Pal Prince Kumar Nandi Manmath Kumar Singh Narendra Kumar Jan Mar 2014 Detoxification of Croton tiglium L seeds by Ayurvedic process of Sodhana Ancient Science of Life 33 3 157 161 doi 10 4103 0257 7941 144619 PMC 4264303 PMID 25538350 External links EditPhorbols at the US National Library of Medicine Medical Subject Headings MeSH Wikimedia Commons has media related to Phorbols Retrieved from https en wikipedia org w index php title Phorbol amp oldid 1119948514, wikipedia, wiki, book, books, library,
