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RU-2861700-C1 - NOVEL MICROBUBBLES WITH PROTEIN-COMPOSITE SHELL FOR ULTRASOUND DIAGNOSTICS AND THERAPY

RU2861700C1RU 2861700 C1RU2861700 C1RU 2861700C1RU-2861700-C1

Abstract

FIELD: pharmaceuticals. SUBSTANCE: group of inventions relates to a composition for use in ultrasound diagnostics, containing gas-filled microbubbles, the shell of which is formed by complexes of albumin with a stabiliser selected from a polyamine, an amino acid or a polymer, and a carrier representing a physiologically acceptable aqueous medium, wherein the gas is air, an inert gas or a fluorine-containing biocompatible gas; the albumin is serum albumin; the polyamine is an oligomer, polymer or pharmaceutically acceptable salt of a compound characterised by the general formula H 2 N–[(CH 2 ) 2-8 –NH] 1-200 –(CH 2 ) 2-8 –NH 2 , having a molecular weight of not more than 15 kDa; the amino acid is tryptophan or tyrosine or a pharmaceutically acceptable salt of said amino acid; the polymer is a biocompatible water-soluble copolymer or a pharmaceutically acceptable salt thereof, with a molecular weight of not more than 15 kDa, characterised by the formula: as well as to a method for producing the above composition. EFFECT: development of compositions containing a new type of microbubbles which combine the advantages of protein and polymer microbubbles, providing improved parameters of stability, biocompatibility and acoustic characteristics; in the development of microbubbles which have an average diameter of 0.5 to 10 mcm, in more preferred embodiments 1-5 mcm, wherein the uniformity of size distribution is not more than 20% deviation from the average diameter, which makes it possible to obtain a uniform ultrasound signal, stable contrast without additional artefacts; in that the density (concentration) of microbubbles in the solution is about 10 8 -10 11 bubbles/ml, which provides good contrast and high stability; in that the developed microbubbles have high stability in physiological solutions: not less than 2 hours at a temperature of 37°C; in that the developed microbubbles also have high storage stability: not less than 6 months at 4°C; in that the developed microbubbles provide a good ultrasound response, have wide functional capabilities and can be used for contrast enhancement in ultrasound diagnostics for various organs and structures, including, but not limited to, targeted imaging of vascular structures, tumours, cavities, including the bladder and female reproductive system, inflammatory processes; in that the developed microbubbles have good biocompatibility and high safety; in that the composition of the shell of the developed microbubbles, represented by modified albumin, provides wide possibilities for their modification for specific medical needs, including the possibility of functionalisation with targeting ligands for targeted delivery, drugs for therapy and theranostics, including for overcoming cellular barriers, including the blood-brain barrier, photo/sonodynamically active drugs for sonodynamic therapy. 12 cl, 13 dwg, 2 ex

Inventors

  • Rudakovskaya Polina Grigorevna
  • Estifeeva Tatyana Mikhajlovna
  • GORIN DMITRIJ ALEKSANDROVICH
  • Barmin Roman Aleksandrovich
  • Nechaeva Anna Mikhajlovna
  • Mezhuev Yaroslav Olegovich

Dates

Publication Date
20260507
Application Date
20250414

Claims (20)

  1. 1. A composition for use in ultrasound diagnostics, containing microbubbles filled with gas, the shell of which is formed by complexes of albumin with a stabilizer selected from a polyamine, an amino acid or a polymer, and a carrier that is a physiologically acceptable aqueous medium, wherein
  2. - the gas is air, an inert gas or a fluorinated biocompatible gas;
  3. - albumin is serum albumin;
  4. - the polyamine is an oligomer, polymer, or pharmaceutically acceptable salt of a compound characterized by the general formula
  5. H 2 N–[(CH 2 ) 2-8 –NH] 1-200 –(CH 2 ) 2-8 –NH 2 , having a molecular weight of no more than 15 kDa;
  6. - the amino acid is tryptophan or tyrosine or a pharmaceutically acceptable salt of the said amino acid;
  7. - the polymer is a biocompatible water-soluble copolymer or its pharmaceutically acceptable salt, with a molecular weight of no more than 15 kDa, characterized by the formula:
  8. in which
  9. R, X and M are chosen independently, with
  10. X denotes the bridging group -S-, -N-, -NH-, -CO-O-, -CO-NH-;
  11. R is
  12. - branched or unbranched -C 6 -C 18 -alkyl-COOH,
  13. - branched or unbranched -C 6 -C 18 -alkyl-OH,
  14. - branched or unbranched -C 6 -C 18 -alkyl-NH 2 ,
  15. - branched or unbranched -C 6 -C 18 -alkyl, optionally substituted with one or two C 6 -aryl groups,
  16. - branched or unbranched –(C j -alkyl)-(C 6 -aryl)-(C z -alkyl),
  17. where j and z are integers from 1 to 17, and the sum of j + z is from 6 to 18;
  18. - branched or unbranched –(C k -alkyl)-(C 6 -aryl)–Y-(C 6 -aryl)-(C m -alkyl),
  19. where Y is a covalent bond or -C p -alkyl, and where k, m and p are integers from 1 to 16, with the sum of k + m + p being from 6 to 18;
  20. - branched or unbranched C 6 -C 18 -alkyl having at least one N atom in the chain, with the formation of a primary, secondary or tertiary amine,

Description

Field of technology The invention relates to the field of biomedical chemistry, materials for use in medical ultrasound diagnostics and therapy, namely to the development of medical contrast agents and agents for therapy using ultrasound. State of the art Several commercial microbubble-based contrast agents for ultrasound diagnostics have been developed and are actively used. These agents are stabilized gas microbubbles that enhance ultrasound imaging by increasing the contrast of vascular structures and tissues. One of the most well-known and widely used contrast agents is SonoVue, a suspension of phospholipid-coated microbubbles filled with sulfur hexafluoride ( SF6 ). This agent is used for ultrasound contrast imaging of blood vessels and organs, providing improved visualization (see, for example, patent RU 2697370 C1, published August 13, 2019). In the United States and some other regions, this agent is known as Lumason for use in cardiac imaging. Disadvantages of this agent include the limited stability of microbubbles in the bloodstream, which reduces the duration of their contrast effect, as well as the lack of the possibility of their targeted delivery or therapeutic functionalization. Another well-known agent is Definity (US Patent No. 10588988 B2, published March 17, 2020), a contrast agent consisting of phospholipid-coated microbubbles filled with octafluoropropane ( C3F8 ). The drug is used for cardiovascular ultrasound imaging. Although the drug is highly stable in the bloodstream, it can cause allergic reactions in some patients. There are also no options for functionalizing the shell for targeted delivery of therapeutic agents. Sonazoid (WO 2020127992 A1, published June 25, 2020) is a microbubbles coated with hydrogenated egg phosphatidylserine and filled with perfluorobutane ( C4F10 ). This product is designed for liver contrast ultrasound imaging and is used in oncology. It is not intended for general ultrasound imaging of vascular structures and internal organs, and does not have the ability to be functionalized for targeted therapy. Optison is an ultrasound contrast agent consisting of albumin-coated microbubbles filled with a perfluorocarbon gas (US Patent 8586005 B2, published November 19, 2013). Unlike other agents, Optison has higher biocompatibility due to the use of a natural protein shell. However, Optison microbubbles have a relatively short circulation time in the bloodstream, limiting the duration of examinations using this contrast agent. Furthermore, the use of unmodified albumin makes the manufacturing process more complex. Despite the fact that a variety of contrast agents with different structures are known, there remains a need for new approaches and methods for obtaining microbubbles that are effective in increasing image contrast during ultrasound diagnostics, are capable of remaining in the bloodstream for a sufficiently long time, and the surface of which can be additionally functionalized. Disclosure of invention The objective of the present invention is to create gas-filled microbubbles suitable for use in medical ultrasound diagnostics and therapy, with improved stability, biocompatibility, and functional properties. This development aims to overcome the shortcomings of existing contrast agents, such as the instability of protein bubbles, the weak ultrasound response of polymer shells, and the rapid degradation of lipid structures. To solve the problem, a composition has been developed for use in ultrasound diagnostics or therapy using ultrasound radiation, containing microbubbles filled with gas, the shell of which is formed by complexes of albumin with a stabilizer selected from a polyamine, amino acid or polymer, and a carrier that is a physiologically acceptable aqueous medium, wherein - the gas is air, an inert gas or a fluorinated biocompatible gas; - albumin is serum albumin; - the polyamine is an oligomer, polymer, or pharmaceutically acceptable salt of a compound characterized by the general formula H 2 N-[(CH 2 ) 2-8 -NH] 1-200 -(CH 2 ) 2-8 -NH 2 , having a molecular weight of no more than 15 kDa; - the amino acid is tryptophan or tyrosine or a pharmaceutically acceptable salt of the said amino acid; - the polymer is a biocompatible water-soluble copolymer or its pharmaceutically acceptable salt, with a molecular weight of no more than 15 kDa, characterized by the formula: , in which R, X and M are chosen independently, while X denotes the bridging group -S-, -N-, -NH-, -CO-O-, -CO-NH-; R is - branched or unbranched -C 6 -C 18 -alkyl-COOH, - branched or unbranched -C 6 -C 18 -alkyl-OH, - branched or unbranched -C 6 -C 18 -alkyl-NH2, - branched or unbranched -C 6 -C 18 -alkyl, optionally substituted with one or two C 6 -aryl groups, - branched or unbranched -(C i -alkyl)-(C 6 -aryl)-(C z alkyl), where j and z are integers from 1 to 17, and the sum of j+z is from 6 to 18; - branched or unbranched -(C k -alkyl)-(C 6 -aryl)-Y-(C 6 -aryl)-(C m -alkyl), where Y