RU2698101C2 - Radiopharmaceutical composition for therapy of inflammatory joint diseases based on radionuclide 188re and human albumin microspheres, as well as a composition and a method for production thereof - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions refers to nuclear medicine. A kit for preparing a suspension of polypeptide biodegradable microparticles for radiosynovectomy in isotonic 0.9 % aqueous solution of sodium chloride labeled with a rhenium isotope consists of auxiliary reagents: antioxidant – ascorbic acid in amount of 7 mg, rhenium reducing agent to lower valence state – stannous chloride dihydrate – 11.4 mg, emulsifier – polysorbate-80 – 1.25 mg, polypeptide carrier of radionuclides – microspheres of human blood albumen with diameter of 5–10 mcm – 5 mg, a transchelator and a pH stabilizer – K,Na-tartrate (tartrate K, Na) – 10 mg, wherein all auxiliary reagents are packaged in three bottles: in bottle No. 1 contains a mixture of a reducing agent and an antioxidant, in bottle No. 2 there is a mixture of a polypeptide carrier of radionuclide and emulsifier atoms, bottle No. 3 contains a transchelator and a pH stabilizer, wherein the contents of each flask are sterile and lyophilised. Also disclosed is a sequential three-stage method of producing a suspension of polypeptide biodegradable microparticles for radiosynovectomy.

EFFECT: group of inventions provides treating inflammatory joint diseases.

4 cl, 4 tbl

Description

Description of the invention:

A radiopharmaceutical composition for the treatment of inflammatory joint diseases based on the ¹⁸⁸ Re radionuclide and human blood albumin microspheres, as well as the composition and method for its preparation.

The present invention relates to nuclear medicine in general and to radiopharmaceutical therapeutic drugs in particular. As an invention, a composition and method for producing a suspension of polypeptide macromolecules with a diameter of 5-10 μm labeled with rhenium radionuclide ( ¹⁸⁸ Re), as well as a method for using the resulting radiopharmaceutical composition for the treatment of rheumatoid joint diseases, will be considered.

For more than four decades, radio synoviortez (RSO) has been a proven important method for the local treatment of chronic inflammatory joint diseases in the context of medical and orthopedic advances. The term radio synoviortez was created by Delbarre et al. [1] and means restoration (orthosis) of the synovial membrane under the influence of ionizing radiation. With intra-articular administration of radiopharmaceuticals for RNO, a radiopharmaceutical preparation (RFP) is localized inside the joint, which has a beneficial effect on the inflammatory processes of the synovial membrane. The therapeutic effect is achieved due to the beneficial effects of ionizing radiation on synovial proliferation, inhibiting its development and leading to further regression followed by fibrous compaction. As a result, a decrease in inflammation in the joint and a decrease in pain.

 In the literature you can find the term "radio synovectomy" - a synonym for the term "radio synoviorthesis."

The first descriptions of the method belong to Ishido (in animals) [2] and Fellinger and Schmid (in relation to people) [3].

With the local use of radioactive substances, an attempt is made to influence the painful destructive process in the joint, which is an alternative to surgical synovectomy. Basically, RNO is intended for topical treatment of almost all types of chronic synovitis. The main indications for RSD are rheumatoid arthritis, seronegative spondylarthropathy, hemophilia with hemophilia, recurrent intraarticular effusion (after arthroscopy), pigmented villonodular synovitis, osteoarthritis, after joint prosthetics and undifferentiated arthritis. A large study was conducted by Kresnik et al. [4] on 2190 treated joints and showed high therapeutic efficacy (73 ± 17)%. The greatest number of responses to treatment was obtained with hemophilic arthritis. In addition to reducing inflammation in arthritis, PCO can lead to a decrease in hypervascularization of the synovial membrane, followed by a significant reduction in bleeding episodes. The response to rheumatoid arthritis was lower and depended on the stage of the disease. Efficiency (73 ± 12)% was recorded in patients with the first stage of synarthrosis according to Steinbrocker classification and with values (52 ± 24)% in patients with the third stage (Table 1). To obtain the best effect, the patient should receive treatment as soon as possible, to prevent the development of arthritis.

Table 1 - Response to treatment with radiosynovectomy of various joints from the study of Kresnik and others.

Type of disease Response to treatment (%) Total number 73 ± 17 Rheumatoid arthritis 65 ± 15 Osteoarthritis 52 ± 15 Hemophilic arthritis 91 ± 4 Stage I RA 73 ± 12 II stage of RA 64 ± 17 III stage of RA 52 ± 24

The most common radiopharmaceuticals used for OCD are: yttrium citrate colloid ( ⁹⁰ Y) for knee joints, rhenium ( ¹⁸⁶ Re) colloid sulfide for medium joints and erbium ( ¹⁶⁹ Er) colloid citrate for small joints with an optimum particle size of 2 microns up to 10 microns [5]. Many different beta-emitting radionuclides were used for labeling colloids and could potentially be used for SAR as part of the radiopharmaceutical (Table 2).

Table 2 - Physical characteristics of the used radioisotopes for radio synovectomy.

Radionuclide β E _max, MeV γ - radiation,% (keV) Mileage in fabric, max, mm Depth of effective passage, min, mm Half life
Days (d)
Hours (h) Applicable activity ^* from literature (MBq) Yttrium – 90 2.26 - 11.0 3.6 2.7 d 185 - 250 Rhenium –186 0.98 9 (137) 3,7 1,2 3.7 d 37 - 185 Erbium – 169 0.34 - 1,0 0.3 9.5 d 15 - 37 Rhenium – 188 2.10 15 (155) 11.0 3.8 17.0 h 555 - 920 Holmium –166 1.84 6 (81) 8.4 3.3 26.9 h 1,111 Dysprosium – 165 1.30 4 (90) 5.7 1.3 2.3 h 9,990 Gold – 198 0.95 95 (412) 3.9 1,0 2.7 d 18 - 370 Samaria – 153 0.81 28 (103) 2.5 0.8 46.8 h 590

It is known from the world prior art to use the above radionuclides, in particular the ¹⁸⁸ Re radionuclide, as an active substance in the composition of the RPF during RNO. Inorganic ¹⁸⁸ Re colloids of various compositions are mainly used for these purposes.

For example, a known method for producing a radiopharmaceutical composition based on the ¹⁸⁸ Re radionuclide in the form of a tin colloid [6]. This patent protects a method for producing a radiopharmaceutical composition from a pre-prepared set of reagents and a solution of sodium perrenate ( ¹⁸⁸ Re) obtained from an ¹⁸⁸ W / ¹⁸⁸ Re generator. The basis of this method is the use of two bottles with auxiliary lyophilized reagents. One of the bottles contains tin dichloride dihydrate and polysorbate-80, which provide the formation of a colloid with the rhenium-188 radionuclide included in its composition. Another bottle contains a pH stabilizer. Sequential mixing of the contents of two vials with the eluate of the ¹⁸⁸ W / ¹⁸⁸ Re generator results in a radiopharmaceutical composition, the active substance of which is colloidal particles containing the ¹⁸⁸ Re radionuclide. The formed particles are in isotonic 0.9% saline. Their size lies in the range from 2 to 10 μm (acceptable particle size for intraarticular injection) with a labeling yield of more than 80%, the radiochemical purity of the drug is more than 90%, the pH of the finished composition is in the range of 3.2 - 5.9. The parameter for the yield of particle labeling in a given range is very important, since particles with sizes less than 2 μm have an increased probability of leaving the joint area, diffusing in the patient’s body, which leads to an increase in unreasonable radiation loads during XRD. But in this case, the formation of particles of the desired size is statistical in nature, since their formation occurs during the reaction from low molecular weight products. In our invention, we will use a narrower range of particles, namely from 5 to 10 microns, which will reduce the likelihood of radioactivity leaving the joint. It will be possible to ensure such precise control by using a polypeptide carrier of a predetermined size, the composition of which will incorporate the ¹⁸⁸ Re radionuclide, which, in turn, will allow for a tagging yield of more than 95%.

 It is possible to use human blood albumin microspheres as such polypeptide macromolecules, the use of which as a carrier of radionuclides in the radiopharmaceutical has been studied for several decades in world practice, primarily due to their affinity with the human body.

The authors of the patent application [7] defend the composition and method of producing a radiopharmaceutical preparation based on rhenium-188 or rhenium-186 radionuclide and human blood albumin microspheres with a diameter of 20 to 40 microns, as well as the possibility of applying the obtained radiopharmaceutical composition for therapeutic purposes unresectable malignant tumors in the liver. The composition is based on the use of three vials with lyophilized auxiliary substances, the successive mixing of which leads to the formation of an injectable form of the drug, which is the basis of the production method protected in the patent application. The formed preparation consists of microspheres of human blood albumin with rhenium-188 ions incorporated into the composition of the polypeptide molecule (obtained from an ¹⁸⁸ W / ¹⁸⁸ Re generator or from a stationary extractor) or rhenium-186 (obtained in a nuclear reactor). The size of the formed particles contributes to the transarterial radioembolization of the capillaries that feed the liver carcinoma cells. Thus, the synergistic effect of reducing the blood supply to tumor cells with local exposure to ionizing radiation leads to targeted cytotoxic effects. This application protects the use of human blood albumin microspheres labeled with ¹⁸⁸ Re radionuclide, as in the present patent application, however, the fundamental difference lies in the composition of the kits for the preparation of the radiopharmaceutical composition, namely in the number of reagents, as well as in the diameter of the labeled microspheres. As a result, the further practical application of the finished RFP differs.

Also known is a method for producing radionuclide-labeled rhenium-188 or rhenium-186 microspheres of human blood serum albumin, where rhenium at the stage suitable for labeling microspheres of albumin is in the form of Re-tricarbonyl ion ( ^186/188 Re- (H ₂ O) ₃ (CO) ₃ ) ⁺ ) [8], the formation of which occurs due to the interaction of ^186/188 ReO ₄ ^- with BH ₃ NH ₃ as a reducing agent in an atmosphere of carbon monoxide. This method is able to complicate the production process of the finished radiopharmaceutical, since it is necessary to carry out work in an atmosphere of carbon monoxide. In the method of obtaining microspheres labeled with radionuclide ¹⁸⁸ Re protected by us, there is no such need.

It is worth noting the patent [9], in which the potential carriers of the ¹⁸⁸ Re radionuclide are not only microspheres of human serum albumin in the blood, but polypeptide molecules in general, in which at least 6% of amino acids contain a sulfhydryl group in the side chain. This protein composition is pretreated with a reducing agent capable of reducing disulfides to sulfhydryls before radiolabeling. According to the claims, a peptide composition containing a radioactive isotope Re-186, Re-188, or a combination of Re-186 and Re-188 is protected as a beta emitter. The specified composition assumes essentially: a spherical shape of a microsphere of a radioisotope labeled protein (peptide) of a spherical shape, used in order to conduct radiation therapy of a mammal. Spherical microspheres contain a radioactive isotope selected from the group consisting of Re-186, Re-188, or a combination of Re-186 and Re-188; these microspheres have a diameter of from 10 to 30 microns. According to the description of the invention, protein - including albumin. However, the size of rhenium-188-labeled microspheres lying in the interval we are protecting is optimal for conducting RNO, since exceeding the diameter of more than 10 μm can increase the complexity of microsphere capture by macrophages of the inner surface of the synovial membrane of the joint.

The objectives of the present invention include the creation of a suspension of polypeptide macromolecules labeled with ¹⁸⁸ Re isotope for nuclear medicine with the elimination of the above disadvantages of existing developments in world practice, with the further use of the resulting radiopharmaceutical composition for the treatment of inflammatory joint diseases.

As a first object of the invention, the composition of the kit for preparing a suspension of polypeptide biodegradable microparticles will be considered. The set for the preparation of the radiopharmaceutical consists of three vials of a drug drip with a capacity of 10 ml, hermetically sealed with rubber stoppers and crimped with aluminum caps. Each of the bottles in the kit is labeled with the number and series. In each bottle, the reagents are in lyophilized form (Table 3). The contents of each vial are sterile.

Table 3. The composition of the set of lyophilisates for the preparation of a suspension of albumin microspheres labeled with a ¹⁸⁸ Re radionuclide with an activity of 37 to 1000 MBq.

Name
lyophilisates Bottle number 1 Bottle number 2 Bottle number 3 Ascorbic
acid mg SnCl ₂ 2H ₂ O,
mg ISA 5-10 microns,
mg Twin 80
mg K, Na tartrate, mg A set of lyophilisates for the manufacture of 7 11,4 five 1.25 10.0

№ 1 vial contains sterile lyophilizate reductant mixture - tin dichloride aqueous 2 (SnCl ₂ 2H ₂ ^O.) And an antioxidant - ascorbic acid (C ₆ H ₈ O _6). Tin dichloride 2 water is a rhenium reducing agent to a lower valence state.

Bottle No. 2 contains a sterile lyophilizate of a mixture of a carrier of atoms of a radionuclide — microspheres of human albumin with a diameter of 5 to 10 microns and emulsifier - polysorbate-80 (C ₆₄ H ₁₂₄ O ₂₆ ). MCAs are spherical monolithic particles with a mirror surface, their density is (1.26 ± 0.12) g / cm3. 5 mg of MSA contains (28,500,000 ± 545,000) units, the average radius of MSA is ~ 7.5 microns. Tween-80 in the composition of the components for the preparation of the suspension is used to improve the wettability of the albumin microspheres, since their surface has hydrophobic properties.

Bottle No. 3 contains a sterile lyophilisate of a transhelator and a pH stabilizer. For these purposes, the salt of tartrate-Na, K (sodium potassium tartrate) is used, which is a ligand that forms a complex with ¹⁸⁸ Re with a low stability constant followed by a religanding property. In addition, sodium potassium tartrate is a reagent that supports the buffering of the reaction mixture and acts as a pH stabilizer.

The next object of protection is a method for preparing the injectable form of the finished radiopharmaceutical based on microspheres of human blood albumin 5-10 microns and rhenium-188, which consists of three stages:

Stage number 1

Isotonic sodium perrenate ( ¹⁸⁸ Re) solution with a volume of 2 to 4 ml and an activity of 37 to 1300 MBq / vial is transferred by puncture of the rubber stopper with a disposable sterile syringe into the vial with sterile lyophilisate No. 1, which is then inserted into a shaker with a rotation speed of 100-150 rpm for 5 minutes. As a result, a solution with a pH from 1.5 to 2.5 is formed in vial No. 1.

Stage number 2

At the end of the previous stage, the solution formed in the bottle No. 1 by puncture of the rubber stopper is taken with a disposable sterile syringe. Once the entire solution has been collected, remove the needle and put on a syringe a membrane filter with a pore diameter of 0.22 μm, install a new needle and filter the contents of the syringe by puncturing the rubber stopper into a vial with sterile lyophilisate No. 2, thus providing additional sterilization of the solution that enters the bottle number 2. Then the bottle number 2 is placed in the oven, where it is heated at a temperature of 90-99 ^about With periodically mixing. The heating time depends on the volume of the heated solution, with a value of 2 ml, the heating time is 1 hour, with 3 ml - an hour and a half, and at 4 ml - 2 hours. This heating interval is explained by a decrease in the concentration of reacting substances, and therefore the need to extend the reaction time to achieve similar results. Upon completion of heating, a suspension of albumin microspheres labeled with rhenium isotope 60-80% is formed in vial No. 2; the pH of the solution is in the range from 1.5 to 2.5.

Stage number 3

At the end of the previous stage, the bottle No. 2 is removed from the oven and allowed to cool at room temperature for 10-20 minutes. The cooled suspension by puncture of a rubber stopper is taken out with a sterile disposable syringe and transferred into a bottle with lyophilisate No. 3, which is then placed in a shaker for 10 minutes at a speed of 100 rpm. After mixing, a suspension of albumin microspheres is formed with a labeling reaction yield of at least 95%, the pH of the supernatant lies in the range from 2.5 to 3.5. The contents of the bottle No. 3 is a ready-made injectable form of the radiopharmaceutical composition.

The result was a suspension of polypeptide macromolecules (MCA) labeled with a ¹⁸⁸ Re radionuclide, the composition of which is given in table. four.

Table 4. The composition of the set of lyophilisates for the preparation of a suspension of albumin microspheres labeled with a ¹⁸⁸ Re radionuclide with an activity of 37 to 1000 MBq.

Rhenium-188 37 - 1000 MBq Albumin Microspheres 1.25–2.5 mg Vitamin C 1.75–3.5 mg Tin dichloride 2 water 2.85–5.7 mg Polysorbate-80 0, 312–0.625 mg Potassium Sodium Tartrate 4-Aqueous 2.5-5.0 mg Sodium chloride 9.0 mg Water for injections 1 ml

The possibility of using a suspension of 5-10 μm albumin microspheres labeled with ¹⁸⁸ Re radionuclide for medical purposes as a potential therapeutic radiopharmaceutical was investigated. MCA labeled with the ¹⁸⁸ Re isotope were studied in laboratory animals (rabbits 6 pcs., Females weighing 1.5 kg on average, and rats 24 pcs. Females weighing 180 g on average) with an artificially created model of acute aseptic synovitis of the knee joint half of the animals from each group. To carry out studies, 0.05 ml of the drug with a volumetric activity of 70.8 MBq / ml was injected intratarticularly into the joint to rats; the dosage for rabbits was 0.1 ml of the drug with a volumetric activity of 192.0 MBq / ml, i.e. 3.6 and 19.2 MBq per animal, respectively.

The data obtained showed that after intraarticular administration of the RFP under study to intact rats, most of the activity (after 3 hours more than 95%, at other time points - more than 97%) remained fixed in the joint for 72 hours of the study. Accumulation in healthy organs and tissues did not exceed 0.5-0.8%. Similar results were obtained for animals with a model of pathology of the knee joint. Thus, we can say that the size of the labeled microspheres is optimal for fixing the source of ionizing radiation in the target organ, namely in the joint. The main source of dose loads on healthy tissues, with a high degree of probability, were unbound ¹⁸⁸ Re ions diffusing from the joint cavity (one of the indicators of the quality of the finished radiopharmaceutical is labeling yield of at least 95%).

The therapeutic efficacy of radiopharmaceuticals based on albumin microspheres 5-10 μm with rhenium-188 in animals with model pathology has also been studied. Studies of the therapeutic efficacy of radiopharmaceuticals "MCA 5-10 μm, ¹⁸⁸ Re" showed the effectiveness of intra-articular administration of radiopharmaceuticals according to criteria for reducing pain sensitivity of the affected limb in rats and the positive dynamics of recovery of motor activity in rabbits for 7 days.

Thus, the protected composition of lyophilized kits for preparation, as well as a method for producing a radiopharmaceutical preparation based on microspheres of human blood albumin and β - The emitting radionuclide rhenium-188 can be considered a potential drug for the treatment of joint diseases. The results of animal studies confirm its functional suitability and safety of use.

Literature:

1. Delbarre F., Cayla J., Menkes C.J., Aignan J., Roucayrol J.C., Ingrand J. La synoviorthèse par les radioisotopes.// Presse Med. Vol. 76, 1045-1050, 1968.

2. Ishido C., Über die Wirkung des Radiothoriums auf die Gelenke.// Strahlentherapie Vol. 15, 537-544, 1923.

3. Fellinger K., Schmid J., Die lokale Behandlung der rheumatischen Erkrankungen.// Wiener Z Inn Med. Vol. 33, 351–363, 1952.

4. Kresnik E., Mikosch P., Clinical outcome of radiosynoviorthesis: a meta-analysis including 2190 treated joints.// Nucl Med Commun Vol. 23, 683–688, 2002.

5. Schneider P., Farahati J., Reiners C., Radiosynovectomy in Rheumatology, Orthopedics, and Hemophilia // J. of Nuc. Med.// Vol. 46, 48S-54S, 2005.

6. Kodina G.E., Malysheva A.O., Klementyeva O.E., Lyamtseva E.A., Taratonenkova N.A., Voronitskaya N.N., Simonenko N.P., Grafskova T.A., Radiopharmaceutical composition for radiosynovectomy and method for its preparation, RF patent No. 2624237.

7. Zverev A.V., Dorovatovsky S.A., Petriev V.M., Skvortsov V.G., Galkin V.N., Kaprin A.D., Radiopharmaceutical drug for the treatment of primary hepatocellular carcinoma and metastatic formations in the liver , as well as the composition and method of its preparation, patent application of the Russian Federation No. 2017135839.

8. Yu Chia-Yu, Lee Te-Wei, Chen Su-Jung, Chen Liang-Cheng, Lin Chen-Hong, Method for making Rhenium-186/188 labeled human serum albumin microspheres and kit for making the same and method for using the kit, US 2013172532.

9. Day D.E., Ehrhard G. J., Zinn K. R., Radiolabeled protein composition and method for radiation synovectomy, US 5403573

Claims (7)

1. A kit for preparing a suspension of polypeptide biodegradable microparticles for radiosynovectomy in an isotonic 0.9% aqueous solution of sodium chloride labeled with rhenium isotope, consisting of auxiliary reagents: antioxidant ascorbic acid in an amount of 7 mg, rhenium reducing agent to a lower valence states - tin chloride dihydrate in an amount of 11.4 mg, an emulsifier - polysorbate-80 in an amount of 1.25 mg, a polypeptide carrier of radionuclides - microspheres of human blood albumin with a diameter of 5-10 microns in 5 mg, transchelator and pH stabilizer - K, Na-tartrate (K, Na tartrate) in an amount of 10 mg, while all auxiliary reagents are packaged in three bottles: bottle No. 1 contains a mixture of a reducing agent and an antioxidant, bottle No. 2 contains a mixture of a polypeptide carrier of atoms of a radionuclide and an emulsifier, vial No. 3 contains a transhelator and a pH stabilizer, while the contents of each vial are sterile and lyophilized. 2. A kit for preparing a suspension of polypeptide biodegradable microparticles in an isotonic 0.9% aqueous solution of sodium chloride labeled with a rhenium isotope according to claim 1, wherein the ¹⁸⁸ Re radionuclide is used as an isotope. 3. A sequential three-stage method for producing a suspension of polypeptide biodegradable microparticles for radiosynovectomy from the kit of claim 1, wherein:

at the first stage, rhenium radionuclide is transferred to vial No. 1 with subsequent mixing of vial No. 1 for 5 minutes at a shaker speed of 100 to 150 rpm, while the pH of the solution in bottle No. 1 at the end of the first stage is in the range from 1 5 to 2.5;

at the second stage, the contents of the bottle No. 1 after the first stage is transferred to the bottle No. 2 through a membrane filter with pores of 0.22 μm, after which the contents of the bottle No. 2 are periodically mixed at a temperature of from 90 to 99 ° C for 60 minutes, if the reaction volume the mixture is 2 ml; 90 min if the volume of the reaction mixture is 3 ml; 120 min if the volume of the reaction mixture is 4 ml; the pH of the solution in vial No. 2 at the end of the second stage is in the range from 1.5 to 2.5, and the yield of the reaction of labeling polymer biodegradable microparticles with a radionuclide belongs to the range from 60 to 80%;

in the third stage, the contents of the bottle No. 2, after the second stage, is cooled to ambient temperature, and then transferred to the bottle No. 3, followed by stirring for 10 minutes at a speed of 50 to 120 rpm, while at the end of the third stage the contents of vial No. 3 is a suspension of biodegradable polymer microparticles labeled with a rhenium-188 radionuclide with a pH in the range from 2 to 5, with a tagging reaction yield of at least 95%. 4. A sequential three-stage method for producing a suspension of polypeptide biodegradable microparticles labeled with a rhenium isotope according to claim 3, wherein the rhenium isotope solution is an isotonic aqueous solution of 0.9% sodium chloride with a volume of 2 to 4 ml containing a ¹⁸⁸ Re radionuclide with a volume activity from 37 MBq / ml to 3.7 GBq / ml.