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WO2025186494A1 - Vésicules extracellulaires à usage clinique dans le traitement de synucléinopathies - Google Patents

Vésicules extracellulaires à usage clinique dans le traitement de synucléinopathies

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Publication number
WO2025186494A1
WO2025186494A1 PCT/ES2025/070114 ES2025070114W WO2025186494A1 WO 2025186494 A1 WO2025186494 A1 WO 2025186494A1 ES 2025070114 W ES2025070114 W ES 2025070114W WO 2025186494 A1 WO2025186494 A1 WO 2025186494A1
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synuclein
evs
shrna
rvg
hdcs
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WO2025186494A8 (fr
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Álvarez Erviti LYDIA
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Fundacion Rioja Salud
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0639—Dendritic cells, e.g. Langherhans cells in the epidermis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A61K48/0075—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the delivery route, e.g. oral, subcutaneous
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs

Definitions

  • Extracellular vesicles for clinical use in the treatment of synucleinopathies Extracellular vesicles for clinical use in the treatment of synucleinopathies
  • the present invention belongs to the technical field of medicine, in particular it relates to extracellular vesicles derived from human dendritic cells and their clinical use in the treatment of synucleinopathies such as Parkinson's disease.
  • Synucleinopathies are neurodegenerative diseases characterized by the abnormal accumulation of ⁇ -synuclein protein aggregates in neurons, nerve fibers, or glial cells.
  • PD Parkinson's disease
  • DLB dementia with Lewy bodies
  • MSA multiple system atrophy
  • Alpha-synuclein is a neuronal protein abundantly expressed in the brain, specifically in presynaptic nerve endings. It constitutes more than 1% of the total protein in the cytosol of brain cells. In synucleinopathies, such as PD, it accumulates excessively within these cells. These deposits are traces of cellular alterations that indicate neuronal dysfunction and eventually lead to neuronal death, a phenomenon responsible for the motor symptoms of the disease.
  • PD is the second most common neurodegenerative disorder, affecting an estimated 10 million people worldwide (L. Hirsch, et al. The incidence of Parkinson's disease: A systematic review and meta-analysis. Neuroepidemiology 46, 292-300 (2016)) and its prevalence is estimated to double by 2040 (E.R. Dorsey, et al. The emerging evidence of the Parkinson pandemic. J Parkinsons Dis. 8, S3-S8 (2016).
  • PD is clinically characterized by the asymmetric and progressive onset of motor symptoms including tremor, rigidity, and bradykinesia, as well as non-motor symptoms in the form of cognition, depression, and autonomic dysfunction (C. Váradi. Clinical features of Parkinson's disease: The evolution of critical symptoms. Biology (Basel).
  • Intracellular a-synuclein which leads to the loss of dopaminergic neurons, occurs in the substantia nigra pars compacta (SNc) (JL Eriksen, et al. Molecular pathogenesis of Parkinson disease. Arch Neurol. 62, 353-7 (2005).
  • Alpha-synuclein has been linked to both sporadic and familial forms of PD, and multiple evidence supports a central role for this protein in the onset and progression of the pathogenesis of this disease (J. Simón-Sánchez, et al. Genome-wide association study reveals genetic risk underlying Parkinson’s disease. Nat. Genet. 41, 1308-1312 (2009); V. M. Lee, et al. Mechanisms of Parkinson’s disease linked to pathological alpha-synuclein: new targets for drug discovery. Neuron 52, 33-38 (2006); J. Y. Li, E., et al. Lewy bodies in grafted neurons in subjects with Parkinson’s disease suggest host-to-graft disease propagation. Nat. Med.
  • RNA small interfering RNA
  • shRNA short hairpin RNA
  • hairping RNA delivered by adeno-associated virus (AAV) (JH Kordower, et al. (2008)) into the SNc, with a decrease in a-synuclein aggregates and motor deficit observed in transgenic or toxin-based PD models.
  • one of the main challenges for gene therapy against ⁇ -synuclein is to design a delivery system or vehicle that crosses the blood-brain barrier, to easily release molecules into the brain after peripheral administration, and modify the disease, altering genetic expression for prolonged periods, given the chronic condition of PD, preferably using vehicles that do not trigger the activation of the immune system, such as AAVs (A. T. Rodger, et al. 2023).
  • the present inventors developed a gene therapy vehicle based on extracellular vesicles (EVs) targeted to the brain by expressing the rabies virus glycoprotein (RVG) peptide on the outer surface of the EVs (RVG-EVs), capable of delivering siRNA and shRNA minicircles (MCs) into the brain after intravenous administration, resulting in gene silencing (L. Alvarez-Erviti, et al. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat. Biotechnol. 29, 341-345 (2011); J. M. Cooper, et al. Systemic exosomal siRNA delivery reduced alpha-synuclein aggregates in brains of transgenic mice. Mov. Disord. 29, 1476-1485 (2014); M. Izco, et al. Systemic exosomal delivery of shRNA minicircles prevents Parkinsonian pathology. Mol. Ther. 27, 2111-2122 (2019).
  • EVs extracellular vesicles
  • MCs Minicircles
  • M. Izco et al. 2019 are double-stranded DNA vectors that contain the transgene expression cassette without additional bacterial sequences (M. Izco et al. 2019), so they are smaller in size. Furthermore, they promote greater transgene expression over longer periods and therefore have the potential to modulate target gene expression for longer periods.
  • ⁇ -synuclein shRNA MCs loaded into RVG-EVs were tested in a model progressive PD in mice, observing its potential to reduce long-term brain ⁇ -synuclein expression, decrease ⁇ -synuclein aggregates, as well as prevent the loss of dopaminergic neurons in the SNc, avoiding motor deficits (/zco et al. 2019).
  • these results are not directly applicable to clinical use in humans, since the application of this technology to clinical trials requires the development of a source of modified human EVs compatible with personalized therapy, since being a chronic and progressive disease the patient will require long-term treatment.
  • FIG. 1 Loading optimization and validation of anti- ⁇ -synuclein shRNA-MC-loaded hRVG-EVs.
  • SH-SY5Y cells overexpressing ⁇ -synuclein were treated with anti- ⁇ -synuclein shRNA-MC via a transfection reaction (TR) or loaded into control hEVs (EV) and hRVG-EVs (RVG-EV).
  • TR transfection reaction
  • EV hEVs
  • RV-EV hRVG-EVs
  • ⁇ -synuclein mRNA (e) and protein (f) levels were quantified and normalized.
  • FIG. 1 Downregulation of ⁇ -synuclein in various brain regions of mice treated with anti- ⁇ -synuclein shRNA-MC delivered by hRVG-EVs.
  • Figure 3 Effect of anti- ⁇ -synuclein shRNA-MC-loaded hRVG-EVs on ⁇ -synuclein pathology in the SNc.
  • Anti- ⁇ -synuclein hRVG-EVs shRNA-MC therapy prevents dopaminergic dysfunction and motor impairments induced by intrastriatal ⁇ -synuclein injections in PFF mice.
  • Dopaminergic innervation in the striatum was quantified by optical density in ipsilateral striatal sections normalized to the contralateral striatum.
  • Figure 7 Proteomic analysis of EVs produced by DCs differentiated in vitro from blood monocytes isolated from control patients, early PD and advanced PD.
  • FIG. 10 Immunofluorescence images of total ⁇ -synuclein in spinal cord (a) and intestine (b) sections from control, PFF, H-EV Syn, H-EV GF, or Ms-EV Syn mice.
  • PD Parkinson's Disease
  • hDCs human dendritic cells
  • an EV derived from hDCs comprises a peptide construct, with sequence SEQ ID NO 1, formed by the fusion of the rabies virus glycoprotein peptide (RVG) and the human Lamp2b protein (hLamp2b) (hereinafter, hLamp2b-RVG) expressed on its surface, where the EV is also loaded with anti- ⁇ -synuclein shRNA minicircles (shRNA-MCs).
  • RVG rabies virus glycoprotein peptide
  • hLamp2b-RVG human Lamp2b protein
  • the use of the EVs of the invention for medical use is contemplated, as well as pharmaceutical compositions or medicaments comprising said EV(s) as an active ingredient.
  • the present invention relates to the use of the EVs of the invention, or of pharmaceutical compositions or medicaments comprising them, in the treatment of synucleinopathies, particularly PD, DLB and MSA.
  • the hLamp2b protein was fused to the RVG peptide.
  • the mRNA encoding the hl_amp2b fusion protein with the RVG tag was synthesized in vitro, resulting in a polynucleotide construct of SEQ ID NO 1, and hDCs were electroporated with said mRNA.
  • the synthesis of the mRNA in c) is carried out using primers of sequence SEQ ID NO 2 and SEQ ID NO 3.
  • mice treated with anti- ⁇ -synuclein shRNA-MCs loaded in human RVG-EVs showed a significant decrease in both mRNA and protein in all 3 regions
  • mice treated with ⁇ -synuclein shRNA-MCs loaded in Ms-RVG-EVs showed a significant decrease in mRNA only in the SNc and cortex, accompanied by a slight decrease in protein levels.
  • Multidose administration of anti- ⁇ -synuclein shRNA-MC loaded into human RVG-EVs does not produce activation of the immune response, implying that this treatment can be administered repeatedly without decreasing its efficacy.
  • the InvitrogenTM mMESSAGE mMACHINETM T7 ULTRA Transcription Kit was used. This kit uses T7 RNA polymerase to synthesize mRNA in vitro, for which it requires a template a linear DNA containing the promoter for the T7 RNA polymerase promoter. The starting point was a plasmid encoding this LAMP2b-RVG sequence with a CMV promoter. To generate the DNA template, a forward primer containing the T7 promoter sequence (SEQ ID NO 2) and a reverse primer (SEQ ID NO 3) were used.
  • SEQ ID NO 2 forward primer containing the T7 promoter sequence
  • SEQ ID NO 3 reverse primer
  • the following POR protocol was used: 95°C 0:12; 56°C 0:12; 68°C 1:30 25x.
  • the mRNA encoding the LAMP2b-RVG protein was then synthesized using the InvitrogenTM mMESSAGE mMACHINETM T7 ULTRA Transcription Kit following the instructions.
  • PBMCs Peripheral blood mononuclear cells
  • 50 ml of PBS was added and PBMCs were separated using a density gradient (Ficoll-Paque PREMIUN 1.073, Cytiva).
  • PBMCs were isolated from blood samples of control individuals and PD patients (15 ml), using a density gradient (Ficoll-Paque PREMIUN 1.073, Cytiva).
  • PBMCs Monocytes were isolated and PBMCs (5 x 10 7 cells per dish, 10 cm dish) were cultured in RPMI-1640 with GlutaMAX (GIBCO-BRL), 10% fetal bovine serum (FCS) without EVs, by centrifugation at 120,000 xg for 60 min and penicillin/streptomycin. Two hours later, the medium was removed and replaced with fresh medium supplemented with 100 ng/ml human granulocyte-monocyte colony-stimulating factor (GM-CSF) (Preprotech) and 20 ng/ml human IL-4 (Preprotech). Seven days later, 106 cells were electroporated with 50 pg of hLamp2b-RVG mRNA (300 V-150 mA).
  • GlutaMAX GlutaMAX
  • FCS fetal bovine serum
  • EVs were harvested by centrifugation at 12,000 xg for 30 minutes to remove cell debris, and the supernatant was centrifuged again at 120,000 xg for 1 h to pellet the EVs. Isolated EVs were resuspended in 0.1 M ammonium acetate using a 27G needle. The size distribution and concentration of EVs were assessed by nanoparticle tracking analysis using an NS500 instrument (Nanosight).
  • EVs were resuspended in PBS and fixed in 2.5% glutaraldehyde solution at 4°C overnight. Samples were dehydrated using a graded ethanol series (30, 50, 70, 80, 90, and 100% * 3) for 10 minutes per step. Samples were coated with gold-palladium by sputter coating, and images were collected using a SEM (model EM-30, COXEM).
  • mice 8–9 weeks old, were purchased from Charles River Laboratories. Mice were housed under standard environmentally controlled conditions with a 12-h light/dark cycle and maintained on an ad libitum food and water diet. Every effort was made to minimize the number of animals used and their suffering.
  • mice received an injection of sonicated murine ⁇ -synuclein preformed fibrils (PFF) into the dorsal striatum and 2 and 45 days later received two intravenous injections of hRVG-EVs containing anti- ⁇ -synuclein shRNA-MCs or anti-GFP shRNA-MCs or two intravenous injections of vehicle (5% glucose).
  • PFF sonicated murine ⁇ -synuclein preformed fibrils
  • mice received an injection of sonicated murine ⁇ -synuclein preformed fibrils (PFF) into the dorsal striatum and 2 and 45 days later received two intravenous injections of hRVG-EVs containing anti- ⁇ -synuclein shRNA-MCs or anti-GFP shRNA-MCs or two intravenous injections of vehicle (5% glucose).
  • Another group of animals injected with PFF ⁇ -synuclein received intravenous injections of mouse RVG-
  • ⁇ -synuclein monomer was assembled into filaments by incubation at 37°C at 5 mg/ml in sterile PBS (pH 7.4) with continuous shaking at 250 rpm for 7 days. Fibrils were pelleted by centrifugation at 10,600 x g for 15 minutes and resuspended in sterile PBS at a concentration of 1 mg/ml. Fibril formation was confirmed by Congo red staining.
  • ⁇ -synuclein PFFs were sonicated for two 6-s cycles at 50% power (10 micron amplitude) using a probe sonicator (Soniprep 150, MSE) immediately prior to surgical injections. A fresh aliquot of sonicated ⁇ -synuclein PFFs was prepared each day of surgery.
  • mice were deeply anesthetized with isoflurane and placed in a stereotaxic frame. Animals were unilaterally injected with 5 pl (two 2.5 pl injections) of freshly sonicated g-synuclein PFF (1 mg/ml) or PBS into the right striatum at a rate of 0.25 pl/min. Striatal coordinates were calculated relative to bregma using the atlas of Paxinos and Watson (Paxinos & Franklin, 2001): AP +0.2 mm, ML -2.0 mm, and DV -3.4 and -2.6 mm. After administration, the needle was left at the injection site for 5 min.
  • RVG-EV containing shRNA-MC 10 EVs and 450 pg of shRNA-MC were electroporated in 10 ml of electroporation buffer (1.15 mM potassium phosphate [pH 7.2], 25 mM KCl, 21% OptiPrep) and electroporated (450 V, 100 mA) in a 4-mm cuvette using a Bio-Rad Gene Pulser Xcell electroporator. Samples were treated with 150 U at 37°C for 30 min. EVs were recovered by ultracentrifugation (120,000 ⁇ g for 1 h) and resuspended in 300 ⁇ l of 5% glucose immediately before intravenous injection into the mouse tail.
  • mice The motor function of the mice was assessed using the wire suspension, negative geotaxis, and limb-hold tests.
  • the wire suspension test was performed before treatment, at 30-day intervals during the study, and before sacrifice, while the limb-hold test was performed on the day of treatment. Sacrifice. Mice were habituated to the testing room for at least 30 minutes before each test and behavioral tests were performed between 09:00 and 12:00 h during the light cycle and were assessed blindly by a trained observer.
  • mice were placed on the wire lid of a conventional cage. The lid was gently shaken three times to ensure the mouse's grip on the bars, and then turned upside down approximately 25 cm above a surface with bedding. The mice's latency to fall off the wire grid was recorded for up to 15 minutes and averaged across two trials (15 minutes apart).
  • mice were held by the midsection of the tail for 10 s.
  • hindlimb position was scored from 0 to 4 according to the following criteria: 0, no limb grasp and normal escape extension; 1, assigned to mice with one hindlimb interlaced and toes showing normal extension; 2, mice with both hindlimbs interlaced inward and toes showing normal extension; 3, both hindlimbs exhibiting a grasp with curled toes and immobility; and 4, mice with the forelimbs and hindlimbs interlaced and crossed with curled toes and immobility (Guyenet SJ, Furrer SA, Damian VM, Baughan TD, La Spada AR, Garden GA. Simple composite phenotype scoring system for evaluating mouse models of cerebellar ataxia. J Vis Exp. 2010 May 21;(39):1787).
  • Serum proinflammatory cytokines including TNF- ⁇ , IFN- ⁇ , IL-1 ⁇ , and IL-6, were measured using the corresponding Proteintech ELISA immunoassay kit according to the manufacturer's protocols.
  • Cell, brain, and spinal cord samples were homogenized in lysis buffer containing 10 mM Tris/HCl (pH 7.4), 0.1% SDS, protease inhibitor cocktail (Thermo Scientific), phosphatase inhibitor cocktail (Thermo Scientific), and DNAase (Promega) and incubated for 1 h at 37°C. The cells were centrifuged at 13,000 rpm for 10 min, and the supernatants were collected. Protein concentrations were determined by the BCA method. Distal intestine samples were homogenized in the same buffer, but containing 8 M urea.
  • Protein samples were solubilized in LDS buffer and reducing agent, separated on 4%-12% Bis-Tris NuPAGE Novex gels (Invitrogen), and transferred to a PVDF membrane. Membranes were blocked with a 10% nonfat dry milk solution prepared in PBS and then incubated with the following primary antibodies: anti- ⁇ -synuclein (Abeam, ref# ab1903), anti-TH (Abeam, ref# ab112), and anti- ⁇ -actin (Abeam, ref# ab6276) antibodies.
  • brain, spinal cord, and intestine samples were collected after perfusion with 4% PFA in PBS, followed by postfixation during immunohistochemistry. night, cryoprotected in 30% sucrose, and frozen. Brains and spinal cords were sectioned into 30 pm coronal sections using a freezing microtome, while intestines were sectioned into 12 pm transverse sections using a cryostat. Sections were washed with TBS, and endogenous peroxidase activity was inactivated by incubation with 3% hydrogen peroxide.
  • the total number of DA neurons in the SNc was assessed by stereology in a series of sections (120 pm intervals between sections) spanning the entire SNc using StereoInvestigator software (MBF Bioscience).
  • the optical density (OD) of TH immunostaining in the striatum was used as an index of the density of striatal dopaminergic innervation.
  • nine representative sections at three coronal levels of the striatum (AP bregma coordinates: +1, 3, +0.5 and -0.4) from each animal were examined and the pixel densities of TH-positive fiber innervation in striatal areas were estimated using ImageJ.
  • the obtained values were normalized by subtracting the OD from the background staining measured in the white matter, where TH-positive innervation is negligible.
  • coronal sections 120 ⁇ m intervals between sections
  • coronal sections 120 ⁇ m intervals between sections
  • the number of phospho- ⁇ -synuclein aggregates was quantified manually at 20x magnification from regularly spaced sections along the rostrocaudal axis of the SNc.
  • Data represent the mean number of aggregates per section.
  • the number of phospho- ⁇ -synuclein aggregates was quantified manually at 20x magnification from regularly spaced sections along the rostrocaudal axis of the SNc.
  • Phospho- ⁇ -synuclein in the cortex and striatum was manually counted on one image per section, with 3 representative sections (AP bregma coordinates: +1, 3, +0.5, and -0.4) from each animal.
  • the number of phospho- ⁇ -synuclein aggregates in the amygdala was manually counted on one image per section, with 2 representative sections (AP bregma coordinates: -1, 9, and -2.2) from each animal.
  • primer sequences were as follows: for ⁇ -synuclein, forward: 5-GCCAAGGAGGGAGTTGTGGCTGC-3' (SEQ ID NO 5); reverse: 5- CTGTTGCCACACCATGCACCACTCC-3' (SEQ ID NO 6); for ⁇ -actin, forward: 5'- TCTACAATGAGCTGCGTGTG-3' (SEQ ID NO 7); reverse: 5'-
  • ⁇ -synuclein expression was normalized to the beta-actin gene.
  • values were calculated using the comparative CT (AACt) method.
  • the hLamp2b protein was fused to the RVG peptide.
  • hLamp2b-RVG mRNA was synthesized in vitro, and hDCs were electroporated with the mRNA; electroporation conditions were optimized using EGFP mRNA.
  • EVs isolated from control hDCs and hDCs electroporated with hLamp2b-RVG mRNA were characterized by NTA (Fig. 1a, 1b) and Western blot (Fig. 1c), and transmission electron microscopy (TEM) (Fig. 1d).
  • hRVG-EVs (3 mg) were loaded with anti- ⁇ -synuclein shRNA-MCs (1 mg) using previously optimized conditions in Ms-RVG-EVs.
  • SH-SY5Y cells overexpressing WT ⁇ -synuclein were treated with anti- ⁇ -synuclein shRNA-MCs using a transfection reagent, or loaded into unmodified hEVs and hRVG-EVs.
  • hRVG-EVs loaded with anti- ⁇ -synuclein shRNA-MCs were intravenously injected into the progressive ⁇ -synuclein PFF mouse model.
  • C57BL6/C3H mice received unilateral intrastinal injections of murine ⁇ -synuclein PFFs.
  • ⁇ -Synuclein protein levels were reduced in the ipsilateral (20% decrease, not significant) and contralateral midbrain (23% decrease, not significant), but were unaffected in the striatum and cerebral cortex of mice treated with anti- ⁇ -synuclein Ms-RVG-EVs shRNA-MC (Fig. 2a-f, Fig. 8).
  • ⁇ -synuclein aggregates in the SNc is associated with decreased survival of dopaminergic neurons.
  • Western blot analysis of TH protein levels in midbrain samples confirmed these data (25% decrease, not significant) (Fig. 9).
  • a similar loss of dopaminergic neurons in the SNc (14% decrease, p 0.049) (Fig.
  • transcriptomic analysis of contralateral cortical samples was performed, the results demonstrated no changes in mRNA expression in PFF mice treated with vehicle, hRVG-EV loaded with anti- ⁇ -synuclein shRNA-MC, or hRVG-EV loaded with anti-GFP shRNA-MC compared to controls (Table 1).
  • proteomic analysis was performed on EVs derived from control individuals, early PD patients, and patients with advanced PD.
  • the proteomic analysis confirmed that EVs produced by DCs differentiated in vitro from blood monocytes isolated from PD patients were suitable as a therapeutic vehicle.
  • a total of 756 proteins were identified, and differential expression analysis showed minimal changes in protein abundance (Fig. 7a, b).
  • the expression of 43 proteins was significantly different in EVs from early PD patients compared to controls, and 20 proteins were significantly different in advanced PD patients compared to controls.
  • GAA-1 lysosomal acid glucosylceramidase-1
  • Fig. 7c a protein associated with familial PD and altered in sporadic PD

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Abstract

La présente invention concerne des vésicules extracellulaires (EV) dérivées de cellules dendritiques humaines (hDC) qui comprennent une construction peptidique RVG-hLAMP, de séquence SEQ ID No 1, exprimée sur sa surface, les vésicules extracellulaires étant chargées avec des mini-cercles de petit ARN en épingle à cheveux (ShRNA-MCs) anti-α-synucléine. L'invention concerne également l'usage clinique desdites vésicules dans le traitement de synucléinopathies. Enfin, la présente invention se rapporte au procédé d'obtention des vésicules extracellulaires à partir de cellues dendritiques humaines (hDC).
PCT/ES2025/070114 2024-03-06 2025-03-05 Vésicules extracellulaires à usage clinique dans le traitement de synucléinopathies Pending WO2025186494A1 (fr)

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ES202430164A ES3035785A1 (es) 2024-03-06 2024-03-06 Vesículas extracelulares para su uso clínico en el tratamiento de sinucleinopatías

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