Kromasil^® notes

Pharmaceutical company uses Kromasil's method development expertise to improve productivity in its purification process

In recent years, India has proven to be an area of strong growth in the peptide market. Numerous Indian companies are developing biosimilars while several others are focusing on research and development (R&D) of novel therapeutic peptides. Issar Pharmaceuticals, located in the Hyderabad, India is an example of a company that has specialized in providing novel therapeutic peptides. The company has successfully synthesized a number of promising peptides in its pipeline, most notably being the Melgain^®, which was commercialized in 2004.

Melgain^® is the commercial name for a decapeptide that can be used as a re-pigmentation agent in the novel treatment of vitiligo. Six years of committed R&D was necessary to identify, refine, and prove the efficacy of this product. While this decapeptide moved through clinical trials on its way to eventual commercialization, several steps were taken to ensure both a cost effective and efficient scale up for production. Within this scope, one crucial step in the production of any peptide is the final purification, known as polishing.

"When designing and scaling up our process, we wanted to use the best equipment and technology possible to guarantee a robust process while keeping the total economy in mind."

The polishing of peptides by HPLC is a step that can be optimized to ensure that the production has the lowest cost possible when looking at a project in the long term. Many factors play a role when determining the purification cost: loadability, media cost, media lifetime, reproducibility, and technical support. Mr. Ram Isanaka, Director at Issar Pharmaceuticals commented, "When designing and scaling up our process, we wanted to use the best equipment and technology possible to guarantee a robust process while keeping the total economy in mind. We decided to use silica from Kromasil to achieve our targets because we believed they were top in class and provided the best long term solution for our purification."

"We decided to use silica from Kromasil to achieve our targets because we believed they were top in class and provided the best long term solution for our purification."

During the development of the purification method for this decapeptide, Issar sent a crude peptide sample to the applications' research lab at Kromasil in Sweden. The expert research chemists at Kromasil used the existing method provided by Issar along with its past experience and knowhow to design a purification method with the total productivity in mind. Loadability is one aspect of the existing method that was greatly improved. Many peptide feed solutions are quite dilute and not conducive for high loading during the purification. This is can be circumvented by starting the method with a low amount of organic solvent (~5-10%), which concentrates the feed at the beginning of the column. Then, the organic solvent concentration can be increased to start elution from the column. One important note is to avoid 100% aqueous conditions in a C18 column to prevent de-wetting. It is important to always keep >4% organic solvent as a minimum.

The improved method was transferred from Kromasil to Issar Pharmaceuticals with great success and showed improved productivity when compared to the previous method. Mr. Isanaka stated, "We were excited and surprised how this method could be scaled up to our 5cm I. D. column and now eventually to our 45cm I. D. column without losing performance. We completed final optimization of the method to suit our latest production improvements and found that the method can be used over and over with high productivity."