International Journal of Dermatology, Vol. 34, No. 1, January 1995 PHARMACOLOGY AND THERAPEUTICS HYDROALCOHOLIC HUMAN PLACENTAL EXTRACT: SKIN PIGMENTING ACTIVITY AND GROSS CHEMICAL COMPOSITION PRAJNAMOY PAL, M.Sc, RABINDRA ROY, PH.D., PIJUSH K. DATTA, M.D., AJIT K. DUTTA, M.D., BANASRI BISWAS, B.Sc, AND R A N J A N B H A D R A , PH.D. Abstract Background. Vitiligo is a pigmentary disorder of the skin of unknown etiology. It is thought to be of autoimmune origin after demonstration of antibody-mediated destruction of melanocytes. Photochemotherapeutic PUVA therapy is widely used in vitiligo with about 33% success. Aqueous or hydroal- coholic extracts of human placenta of ill-defined composition have also been used therapeutically for vitiligo. A hydroalco- holic human placental extract has been developed by us with pigmenting activity based on experimental therapies. Its chemical analysis was the primary objective of this study. Methods. For the guinea pig experiment, 20 drops of the ex- tract or vehicle (60% alcohol) as control was topically applied around the nipples covering the areola zones of male imma- ture white guinea pigs (wt. 175-250 g) daily for 60 days with 15 minutes infrared (IR) exposure used for vascular dilatation and enhancement of the absorption of the extract. Standard methods have been followed for all chemical analyses. Results. The guinea pig experiment showed clear pigmen- tation and hypertrophy of the experimental nipples to vary- ing degrees. Chemical analysis of the extract revealed the presence of small-molecular-weight proteins/peptides, lipids (including glycosphingolipids), carbohydrates, sialic acids, cholesterol, triglycerides, high density lipoproteins (HDL), and others, including amino acids, nucleotides, carotenes, vitamins, etc. Conclusion. Glycosphingolipids, known modulators of B and T cells, were reported capable of inducing adhesion, spreading, and motility of melanoma. It is present in the ex- tract and, therefore, may lead to skin pigmentation through From the Department of Cellular Biochemistry, Indian Insti- tute of Chemical Biology, Calcutta, India; the Department of Dermatology, Medical College, Calcutta, India; and the Der- matology Section, Institute of Child Health, Calcutta, India. Current address (Dr. Roy): Sealy Center for Molecular Sci- ence, University of Texas, Medical Branch, at Galveston, Brakenridge, Calveston, Texas. Address for correspondence: Ranjan Bhadra, Ph.D., Cellular Biochemistry Department, Indian Institute of Chemical Biology, 4, Raja S.C. Mullick Road, Jadavpur, Calcutta 700032, India. induction of melanocytes. Endothelin, a 21-amino acid pep- tide, detected in human placenta and possibly extractable by our process, has been reported to be indispensable for melanocyte growth. IntJ Dermatol 1995; 34:61-66 In recent studies,' it has been demonstrated that the antibody-mediated destruction of melanocytes is one of the prime factors behind vitiligo, a pigmentary dis- order of the skin, affecting 1% of the world popula- tion. No satisfactory therapy is available for this con- dition. Loss of melanocytes is described as the basis of developing macules and patches of depigmented areas in the skin with various distribution and degrees. Among the pathophysiologic disorder relating to the dysfunction and loss of melanocytes or development of vitiligo, a faulty immune response is being considered to be most likely in current studies.'"^ Apart from this, there are other hypotheses such as neural, free-radical, and melanocyte growth factors reduction.-' The current recommended standard therapy for vi- tiligo is the topical or systemic application of psoralens with ultraviolet light commonly known as PUVA thera- py; however, only one-third of the patietits respotid to this therapy*" and it is highly phototoxic.'' Other nonpsoralen photo- and nonphotochemotherapeutic medications, such as phenylalanine, khellin, cortico- steroids, 5-fluorouracil,''''* and aqueous and alcoholic extracts of human placenta'' have also been used for the treatment of vitiligo. An aqueous extract of human placenta possesses the keratinocyte growth factor,^ stimulators of endothelial cell growth,'" and probably the fibroblast growth fac- tor.' Thus, human placental components with growth- promoting activity on skin cells are well documented. Probably based on these observations, the aqueous and alcoholic extracts of human placenta have been used as an experimental therapy in various disorders such as vitiligo;'' however, these types of extracts require sys- temic analysis to identify the active principle and to lay a scientific foundation on a sound basis. One such preparation, claimed to be effective in vitiligo, of un- known composition,*^-" was controversial regarding its 61