Insights into the Mechanism of Action of Highly Diluted Biologics

Abstract

The therapeutic use of Abs in cancer, autoimmunity, transplantation, and other fields is among the major biopharmaceutical advances of the 20th century. Broader use of Ab-based drugs is constrained because of their high production costs and frequent side effects. One promising approach to overcome these limitations is the use of highly diluted Abs, which are produced by gradual reduction of an Ab concentration to an extremely low level. This technology was used to create a group of drugs for the treatment of various diseases, depending on the specificity of the used Abs. Highly diluted Abs to IFN-g (hd-anti-IFN-g) have been demonstrated to be efficacious against influenza and other respiratory infections in a variety of preclinical and clinical studies. In the current study, we provide evidence for a possible mechanism of action of hd-anti-IFN-g. Using high-resolution solution nuclear magnetic resonance spectroscopy, we show that the drug induced conformational changes in the IFN-g molecule. Chemical shift changes occurred in the amino acids located primarily at the dimer interface and at the C-terminal region of IFN-g. These molecular changes could be crucial for the function of the protein, as evidenced by an observed hd-anti-IFN-g-induced increase in the specific binding of IFN-g to its receptor in U937 cells, enhanced induced production of IFN-g in human PBMC culture, and increased survival of influenza A-infected mice.

New insights into the microscopic interactions associated with the physical mechanism of action of highly diluted biologics

“…We have also conducted a series of molecular dynamics (MD simulations) on crystal structures of antibody and antigen complexes with the aim of providing a basis for the interpretation of our experimental measurements. The interpretation of the results obtained is consistent with the data of previously performed structural studies of the action of HD proteins on the conformation of IFN-g 7 , and for the first time explains the physical features of the molecular mechanism of the modifying action of HD samples.…”

Analysis of Emission Infrared Spectra of Protein Solutions in Low Concentrations

“…Above all, on average, the control spectrum is more intensive compared to the other three spectra. Figure 2B shows paired comparisons of the test solutions with the control solution in terms of their emission spectra according to formula (2).…”

“…In this paper, the developed method was used for the study of water protein solutions (interferon-gamma (IFNγ), antibodies to interferon-gamma (Ab to IFNγ)) and glycine buffer (GB) at low concentration. The issue of sensitivity when dealing with low concentrations is of utmost importance, in particular, given the fact that some studies [2][3][4][5][6][7] have revealed the effects observed in protein solutions at low concentrations that can hardly be explained directly by the properties of protein and the solvent, i.e., water. Furthermore, the present study analyzed the properties of the protein (IFNγ) dissolved in the above-mentioned solutions at low concentration.…”

“…(A) Emission spectra of the IFNγ 1 mg/ml solutions supplemented with the 10v% test samples or the control. (B) Paired comparisons of emission spectra of the IFNγ 1 mg/ml solutions supplemented with the 10v% test samples for detecting statistically significant differences from the IFNγ 1 mg/ml solution supplemented with the control according to formula(2).…”

IVIG ameliorate inflammation in collagen-induced arthritis: projection for IVIG therapy in rheumatoid arthritis

“…Despite the significant progress that has been achieved with the administration of biological therapies in changing the natural history of RA, such medications induce immune suppression, which is non‐selective to the pathogenesis of the disease, resulting in higher rates of common and opportunistic infections. Thus, different strategies are used to overcome these issues; for example, processing of antibodies under specific high‐dilution technology to produce drugs that change the conformation state of the antigen and have specific target‐modification activity [8,9], or produce domain antibodies such as nanobodies, recently approved by the Food and Drug Administration (FDA) for the first time [10]. Another highly promising approach is the usage of intravenous immunoglobulins (IVIG) as a therapy for RA.…”