Discovery and development of first in class antifungal caspofungin (CANCIDAS®)--a case study.

Covering: 1985 to 2001.This paper describes a fifteen year journey from concept to clinical discovery and development of the first in class caspofungin acetate (CANCIDAS®) a parenteral antifungal agent. Caspofungin is a semisynthetic derivative of pneumocandin B0, a naturally occurring, lipophilic cyclic peptide isolated from the fungus, Glarea lozoyensis. While the echinocandins had been previously studied for antifungal activity by several organizations, the class was dropped for a variety of reasons. Merck subsequently initiated a research program leading to the discovery and development of caspofungin. The multitude of challenges that ensued during the discovery and development process and which were successfully resolved by multi-disciplinary teams constitute the content of this article. The article consists of five sections that describe the discovery and development of caspofungin in chronological order: (i) discovery of the natural product pneumocandin B0 from fungal fermentations, (ii) fermentation development to improve the titer of pneumocandin B0 to make it commercially viable, (iii) semisynthetic modification by medicinal chemistry to successfully improve the properties of pneumocandin B0 leading to the discovery of caspofungin, (iv) development of commercial semisynthesis and purification and formulation development to improve stability and (v) clinical development and approval of CANCIDAS® as an antifungal drug which subsequently saved thousands of lives.

Nutritional and engineering aspects of microbial process development.

Today we use many drugs produced by microorganisms. However, when these drugs were discovered it was found that the yields were low and a substantial effort had to be put in to develop commercially viable processes. A key part of this endeavor was the studies of the nutritional and the engineering parameters. In this chapter, the basic principles of optimizing the nutritional and engineering aspect of the production process are described with appropriate examples. It was found that two critical components of nutritional medium, carbon and nitrogen source regulated the synthesis of the compounds of interest. Rapidly utilizable carbon source such as glucose supported the growth but led to catabolite repression and alternative carbon sources or methods of addition had to be devised. Inorganic nitrogen sources led to undesirable changes in pH of the medium. Organic nitrogen sources could influence the yields positively or negatively and had to be chosen carefully. Essential nutrients like phosphates often inhibited the synthesis and its concentration had to be maintained below the inhibitory levels. On many occasions, trace nutrients like metal ions and vitamins were found to be critical for good production. Temperature and pH were important environmental variables and their optimum values had to be determined. The media were designed and optimized initially with 'one variable at a time' approach and later with experimental design based on statistics. The latter approach is preferred because it is economical, considers interactions between medium components and allows rapid optimization of the process. The engineering aspects like aeration, agitation, medium sterilization, heat transfer, process monitoring and control, become critical as the process is scaled-up to the production size. Aeration and agitation are probably the most important variables. In many processes dissolved oxygen concentration had to be maintained above a critical value to obtain the best yields. The rheological properties of fermentation broth significantly affect the aeration and mixing efficiency. The removal of heat from the large fermentors can be difficult under certain conditions. However, new designs of impellers, availability of sensors to monitor important physiological and process variables and advent of computers have facilitated successful scale-up of fermentation processes.