PEPTIDOTS Technology

A unique non-denaturing process for protein encapsulation

Drug delivery can offer an injectable product at a competitive edge in several ways. Delivery technologies that result in a more stable plasma drug concentration can improve the efficacy and safety profile of a drug. If a therapeutic protein can be offered as a less frequent or more patient-friendly formulation or device, then there is a significant higher chance that patients will follow the dosing regimen, leading to improved treatment outcomes.

Reducing the number and frequency of administrations importantly increases patient compliance, especially when treating chronic diseases.

Over recent years, there has been a considerable increase in the number of biological pharmaceutical products, mostly in the field of cancer, cardiovascular diseases, vaccination, diabetes, the treatment of angiogenesis disorders, and immunotherapies. One of the major hurdle limiting the use of therapeutic proteins relates to their limited stability, degradation or denaturation upon various physiological and formulation process parameters (enzymatic degradation, pH drops, temperature changes, presence of organic solvents). Moreover, clinical administration of proteins at therapeutically effective doses is difficult, because of their undesirable physicochemical properties in solution, such as aggregation and high viscosity.

In order to address these drawbacks, CARLINA Technologies has developed an innovative technology using a patented nanoprecipitation procedure that allows therapeutic proteins to be stabilized before being inserted into polymeric matrix-based systems (microspheres, gels, implants, etc.)  for controlled-delivery.

To our knowledge, no other method described to date provides a comparably efficient non-denaturing process to encapsulate therapeutic proteins with no loss of protein activity.

How does it work?

PEPTIDOTS is a two-step process. It first consists in a fully rerversible nanoprecipitation step allowing the protein to be concentrated while maintaining its biological activity. In a second stage, the nanoprecipitates are inserted in various types of sustained-delivery systems. This reversible nanoprecipitation step is covered by our IP and represents a brakethrough benefit. Should this step not be performed, most of the released protein would be denaturated therefore becoming inactive.

Our PEPTIDOTS technology stabilizes the protein in solution by adding a non-solvent to the protein suspension. As non-solvents are small hydrophilic molecules now in excess, they replace water molecules leading to a non-denaturing aggregation of proteins assembled into nanoparticles that are ranging between 50 nm and 100 nm in mean diameter. This process is called nanoprecipitation and is driven by a desolvatation of the protein in solution. The excess of the non-solvent promotes protein-protein interactions leading to a thermodynamic equilibrium state under the form of protein nanoparticles (nanoprecipitates). Additional additives can contribute to both increase the efficiency of nanoprecipitation up to 100% or near, and favour protein release from nanoprecipitates-loaded polymeric matrices.

Procédé de nanoprécipitation

In a second stage, the nanoprecipitates are encapsulated, or simply inserted, in various types of sustained-delivery systems including microspheres, gels or implants without been denaturated. Upon administration (i.e. subcutaneously) the polymer degradation forming the encapsulating system will slowly release the protein nanoprecipitates over time. While nanoprecipitates are released in a physiological aqueous environment, proteins will reversibly go back to their initial conformation without being degraded.

Once administered to patients, the nanoprecipitate-loaded matrix made of a biodegradable polymer (such as Poly(Lactic-co-Glycolic) acid (PLGA)) will degrade over time, therefore slowly releasing its content. Once release of nanoprecipitates occurs, the water molecules from physiological fluids are now in excess as compared to non-solvent molecules. Non-solvent molecules in interaction with the protein are now displaced by water molecules. The protein then reversibly comes back from its nanoprecipitate form to its initial native conformation.

We demonstrated that PEPTIDOTS can be used to highly concentrate proteins, including antibodies, in suspensions. High concentration liquid formulations and enhanced stability formulations are key development areas especially in the field of monoclonal antibodies. Either encapsulated or naked nanoprecipitates of proteins can be highly concentrated in non-viscous injectable suspensions. Moreover, highly concentrated suspensions of protein nanoprecipitates allow to obtain very high efficiencies of encapsulation in polymetric matrices (> 80 %).

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  • Non viscous solutions of highly concentrated proteins
  • Release from days to weeks
  • Simple, cost-effective, upscalable manufacturing process
  • Non toxic excipients and polymers used: high development success rate
  • No protein modification: native protein retaining full biological activity
  • Applicable to a large variety of proteins

Learn more about CARLINA Technologies