Abstract
In the continued absence of an effective anti-HIV vaccine, approximately 2 million new HIV infections occur every year, with over 95% of these in developing countries. Calls have been made for the development of anti-HIV drugs that can be formulated for topical use to prevent HIV transmission during sexual intercourse. Because these drugs are principally destined for use in low-resource regions, achieving production costs that are as low as possible is an absolute requirement. 5P12-RANTES, an analog of the human chemokine protein RANTES/CCL5, is a highly potent HIV entry inhibitor which acts by achieving potent blockade of the principal HIV coreceptor, CCR5. Here we describe the development and optimization of a scalable low-cost production process for 5P12-RANTES based on expression in Pichia pastoris. At pilot (150 L) scale, this cGMP compliant process yielded 30 g of clinical grade 5P12-RANTES. As well as providing sufficient material for the first stage of clinical development, this process represents an important step towards achieving production of 5P12-RANTES at a cost and scale appropriate to meet needs for topical HIV prevention worldwide.
Highlights
There is an urgent need for new products that can be used to stem the spread of HIV/AIDS, in developing countries, where over 95% of the estimated 2 million new infections per year
In order to identify an appropriate recombinant expression system for large scale low cost production of 5P12-RANTES, we focused on yeast secretory expression, which has been shown to be robust and effective for other chemokines [14e16], including analogs of RANTES/CCL5 [17,18]
Zeocin-resistant clones obtained by transforming P. pastoris strain Bg08 with the pJAZaMF encoding 5P12-RANTES were screened for 5P12-RANTES expression
Summary
There is an urgent need for new products that can be used to stem the spread of HIV/AIDS, in developing countries, where over 95% of the estimated 2 million new infections per year occur [1]. Among the strategies being explored are those centered on the use of topically applied anti-HIV drugs, which can be used to prevent transmission of the virus during sexual intercourse [2]. 5P12-RANTES [7] is a next-generation fully recombinant analog which was developed using a phage display approach [10]. In addition to having potency and efficacy comparable to that of PSCRANTES [7,11], 5P12-RANTES shows promising stability under storage at elevated temperature and when exposed to vaginal pH, human cervicovaginal lavage and human semen [12]. It has been shown to present a formidable barrier to the generation of escape mutants [13]. 5P12-RANTES represents a highly promising candidate for development as a topical agent for HIV-prevention
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have