Neuro10-9 Pharma, Inc. | 3110 Edwards Mill Rd #300, Raleigh, NC 27612 | 919-923-7401 | ©2015
NEURONANO’s strategy is to develop collaborations with discovery and development companies to enhance the success of their product lines, while bringing its own products forward into clinical studies.
— Sasha Kabanov
NEURONANO Pharma is a biopharmaceutical company developing solutions to some of the most vexing drug development problems facing pharmaceutical companies worldwide today.
NEURONANO has perfected a series of formulation and delivery technologies that when used with current or newly discovered compounds can 1) preserve even very fragile biologics as they traverse the body, 2) improve the transport of polypeptides, proteins, hormones, enzymes and small molecules across cell membranes and tissue barriers, including the blood brain barrier, and 3) improve the release of drugs encapsulated in liposomes.
The Company has proof of concept data in animal systems for hormones, proteins, enzymes and small molecules with therapeutic applications in CNS disorders, metabolic diseases, cancer and ophthalmic disease.
nanoZYME™ is a patent-pending platform technology enabling effective delivery of protein therapeutics to tissues and organs that have proven difficult to access. Our technology incorporates proteins into nano-sized biocompatible polymer coatings through a readily scalable self-assembly process. The platform preserves even very fragile proteins in biological milieu and allows their site-specific delivery across cellular and mucus barriers. The platform has demonstrated in vivo proof of concept across a broad range of proteins that 1) were delivered to the brain across the blood brain barrier, 2) were transported with macrophages to sites of inflammation, 3) crossed the cornea into the eye, and 4) were deposited into lungs through the airways.
poloMAC™ and poxoMAC™ are patented platform technologies for polypeptide delivery based on conjugation of polypeptides with block copolymers to increase polypeptide circulation time and stability, while preserving native confirmation and potency. Compared to commonly used PEGylation systems, these platforms greatly improve transport of non-permeable polypeptides into cells and across diverse biological barriers. Both poloMAC™ and poxoMAC™ have been demonstrated to increase the bioavailability of enzymes and hormones in select brain regions after systemic and nasal administration in animal models.
lipoXOL™ is a patent-pending platform technology that provides a simple way to increase the efficacy of liposomal drugs by improving the release of these drugs from liposomes at the target site of action. Although liposomal formulations of active pharmaceutical ingredients (API) have been previously shown to provide enhanced half-lives and lower side effects compared to API without liposomal formulation, drug entrapment within liposomes can make such formulations less efficacious. While maintaining the superior PK and safety profiles of liposomal formulations, lipoXOL™ technology uses “generally regarded as safe (GRAS)” excipients to enhance the release of API from liposomes within tumors, thus enhancing efficacy at the target site of action.
Technology marvels creation, in modern world, requires freedom of entrepreneurship and its protection by law. Simple, obvious, priceless!
— Dr. Kabanov, Lead Inventor
NEURONANO is actively working on protecting our delivery platforms, technologies, products, compositions, their methods of use, and the processes for their manufacture. At its founding, NEURONANO in-licensed a portfolio of over 30 groundbreaking inventions on an exclusive and worldwide basis from University of Nebraska Medical Center, St. Louis University, University of Washington, and University of North Carolina at Chapel Hill. Following initial licensing, we continue to follow a strategy of filing aggressively on our cutting edge technologies in order to maintain exclusivity of our delivery advantages.
Compositions for Protein Delivery and Methods of Use Thereof
Alexander V. Kabanov, Tatiana Bronich, Elena Batrakova, Howard Gendelman
Compositions and Methods for the Treatment of Lung Inflammation
Alexander V. Kabanov, Devika Soundara Manickam
Nanozyme Compositions and Methods of Synthesis and Use Thereof
Alexander V. Kabanov, Davika Soundara Manickam, Anna M. Brynskikh
Amphiphilic Polymer-Protein Conjugates and Methods of Use Thereof
Alexander V. Kabanov, Xiang Yi, Serguel V. Vinogradov, William A. Banks
Delivery of biotherapeutics to the brain
Xiang YI, Alexander V. Kabanov, William Banks
Amphiphilic polymer-protein conjugates and methods of use thereof
Elena V. Batrakova, Serguei V. Vinogradov, Alexander V. Kabanov
Protein-poly(2-oxazoline) conjugates for enhanced cellular delivery and transport across biological barriers
Alexander V. Kabanov, Jing Tong
Compositions and Methods for the Treatment of Cancer
Daria Y. Alakhova, Alexander V. Kabanov, Yi Zhao
Water Soluble Fullerene Formulations and Methods of Use Thereof
Alexander V. Kabanov, Jing Tong
Yi, X., Manickam, D.S., Brynskikh, A., Kabanov, A.V.
A variety of therapeutic proteins have shown potential to treat central nervous system (CNS) disorders. Challenge to deliver these protein molecules to the brain is well known. Proteins administered through parenteral routes are often excluded from the brain because of their poor bioavailability and the existence of the blood-brain barrier (BBB). Barriers also exist to proteins administered through non-parenteral routes that bypass the BBB. Several strategies have shown promise in delivering proteins to the brain. This review, first, describes the physiology and pathology of the BBB that underscore the rationale and needs of each strategy to be applied. Second, major classes of protein therapeutics along with some key factors that affect their delivery outcomes are presented. Third, different routes of protein administration (parenteral, central intracerebroventricular and intraparenchymal, intranasal and intrathecal) are discussed along with key barriers to CNS delivery associated with each route. Finally, current delivery strategies involving chemical modification of proteins and use of particle-based carriers are overviewed using examples from literature and our own work. Whereas most of these studies are in the early stage, some provide proof of mechanism of increased protein delivery to the brain in relevant models of CNS diseases, while in few cases proof of concept had been attained in clinical studies. This review will be useful to broad audience of students, academicians and industry professionals who consider critical issues of protein delivery to the brain and aim developing and studying effective brain delivery systems for protein therapeutics.
Price, T.O., Farr, S.A., Yi, X., Vinogradov, S., Batrakova, E.V., Banks, W.A., Kabanov, A.V.
Leptin is a peptide hormone produced primarily by adipose tissue that acts as a major regulator of food intake and energy homeostasis. Impaired transport of leptin across the blood-brain barrier (BBB) contributes to leptin resistance, which is a cause of obesity. Leptin as a candidate for the treatment of this obesity is limited because of the short half-life in circulation and the decreased BBB transport that arises in obesity. Chemical modification of polypeptides with amphiphilic poly(ethylene oxide)-poly(propylene oxide) block copolymers (Pluronic) is a promising technology to improve efficiency of delivery of polypeptides to the brain. In the present study, we determined the effects of Pluronic P85 (P85) with intermediate hydrophilic-lipophilic balance conjugated with leptin via a degradable SS bond [leptin(ss)-P85] on food intake, clearance, stability, and BBB uptake. The leptin(ss)-P85 exhibited biological activity when injected intracerebroventricularly after overnight food deprivation and 125I-leptin(ss)-P85 was stable in blood, with a half-time clearance of 32.3 min (versus 5.46 min for leptin). 125I-Leptin(ss)-P85 crossed the BBB [blood-to-brain unidirectional influx rate (K(i)) = 0.272 +/- 0.037 microl/g x min] by a nonsaturable mechanism unrelated to the leptin transporter. Capillary depletion showed that most of the 125I-leptin(ss)-P85 taken up by the brain reached the brain parenchyma. Food intake was reduced when 3 mg of leptin(ss)-P85 was administered via tail vein in normal body weight mice [0-30 min, p < 0.0005; 0-2 h, p < 0.001]. These studies show that the structure based Pluronic modification of leptin increased metabolic stability, reduced food intake, and allowed BBB penetration by a mechanism-independent BBB leptin transporter.
Yi, X., Yuan, D., Farr, S.A., Banks, W.A., Poon, C.D., Kabanov, A.V.
Modification of hydrophilic proteins with amphiphilic block copolymers capable of crossing cell membranes is a new strategy to improve protein delivery to the brain. Leptin, a candidate for the treatment of epidemic obesity, has failed in part because of impairment in its transport across the blood-brain barrier (BBB) that develops with obesity. We posit that modification of leptin with poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide), Pluronic P85 (P85) might permit this protein to penetrate the BBB independently of its transporter, thereby overcoming peripheral leptin resistance. Here we report that peripherally administered leptin-P85 conjugates exhibit biological activity by reducing food intake in mouse models of obesity (ob/ob, and diet-induced obese mouse). We further generated two new leptin-P85 conjugates: one, Lep(ss)-P85(L), containing one P85 chain and another, Lep(ss)-P85(H), containing multiple P85 chains. We report data on their purification, analytical characterization, peripheral and brain pharmacokinetics (PK). Lep(ss)-P85(L) crosses the BBB using the leptin transporter, and exhibits improved peripheral PK along with increased accumulation in the brain compared to unmodified leptin. Lep(ss)-P85(H) also has improved peripheral PK but in a striking difference to the first conjugate penetrates the BBB independently of the leptin transporter via a non-saturable mechanism. The results demonstrate that leptin analogs can be developed through chemical modification of the native leptin with P85 to overcome leptin resistance at the level of the BBB, thus improving the potential for the treatment of obesity.
Tong, J., Yi, X., Luxenhofer, R., Banks, W.A., Jordan, R., Zimmerman, M.C., Kabanov, A.V.
Superoxide dismutase 1 (SOD1) efficiently catalyzes dismutation of superoxide, but its poor delivery to the target sites in the body, such as brain, hinders its use as a therapeutic agent for superoxide-associated disorders. Here to enhance the delivery of SOD1 across the blood-brain barrier (BBB) and in neurons the enzyme was conjugated with poly(2-oxazoline) (POx) block copolymers, P(MeOx-b-BuOx) or P(EtOx-b-BuOx), composed of (1) hydrophilic 2-methyl-2-oxazoline (MeOx) or 2-ethyl-2-oxazoline (EtOx) and (2) hydrophobic 2-butyl-2-oxazoline (BuOx) repeating units. The conjugates contained from 2 to 3 POx chains joining the protein amino groups via cleavable -(ss)- or noncleavable -(cc)- linkers at the BuOx block terminus. They retained 30% to 50% of initial SOD1 activity, were conformationally and thermally stable, and assembled in 8 or 20 nm aggregates in aqueous solution. They had little if any toxicity to CATH.a neurons and displayed enhanced uptake in these neurons as compared to native or PEGylated SOD1. Of the two conjugates, SOD1-(cc)-P(MeOx-b-BuOx) and SOD1-(cc)-P(EtOx-b-BuOx), compared, the latter was entering cells 4 to 7 times faster and at 6 h colocalized predominantly with endoplasmic reticulum (41 ± 3%) and mitochondria (21 ± 2%). Colocalization with endocytosis markers and pathway inhibition assays suggested that it was internalized through lipid raft/caveolae, also employed by the P(EtOx-b-BuOx) copolymer. The SOD activity in cell lysates and ability to attenuate angiotensin II (Ang II)-induced superoxide in live cells were increased for this conjugate compared to SOD1 and PEG-SOD1. Studies in mice showed that SOD1-POx had ca. 1.75 times longer half-life in blood than native SOD1 (28.4 vs 15.9 min) and after iv administration penetrated the BBB significantly faster than albumin to accumulate in brain parenchyma. The conjugate maintained high stability both in serum and in brain (77% vs 84% at 1 h postinjection). Its amount taken up by the brain reached a maximum value of 0.08% ID/g (percent of the injected dose taken up per gram of brain) 4 h postinjection. The entry of SOD1-(cc)-P(EtOx-b-BuOx) to the brain was mediated by a nonsaturable mechanism. Altogether, SOD1-POx conjugates are promising candidates as macromolecular antioxidant therapies for superoxide-associated diseases such as Ang II-induced neurocardiovascular diseases.
Manickam, D.S., Brynskikh, A.M., Kopanic, J.L., Sorgen, P.L., Klyachko, N.L., Batrakova E.V., Bronich T.K., Kabanov A.V.
Development of well-defined nanomedicines is critical for their successful clinical translation. A simple synthesis and purification procedure is established for chemically cross-linked polyion complexes of Cu/Zn superoxide dismutase (SOD1) or catalase with a cationic block copolymer, methoxy-poly(ethylene glycol)-block-poly(L-lysine hydrochloride) (PEG-pLL₅₀). Such complexes, termed cross-linked nanozymes (cl-nanozymes) retain catalytic activity and have narrow size distribution. Moreover, their cytotoxicity is decreased compared to non-cross-linked complexes due to suppression of release of the free block copolymer. SOD1 cl-nanozymes exhibit prolonged ability to scavenge experimentally induced reactive oxygen species (ROS) in cultured brain microvessel endothelial cells and central neurons. In vivo they decrease ischemia/reperfusion-induced tissue injury and improve sensorimotor functions in a rat middle cerebral artery occlusion (MCAO) model after a single intravenous (i.v.) injection. Altogether, well-defined cl-nanozymes are promising modalities for attenuation of oxidative stress after brain injury.
Savalia, K., Manickam, D.S., Rosenbaugh, E.G., Tian, J., Ahmad, I.M., Kabanov, A.V., Zimmerman, M.C.
Excessive production of superoxide (O2(-)) in the central nervous system has been widely implicated in the pathogenesis of cardiovascular diseases, including chronic heart failure and hypertension. In an attempt to overcome the failed therapeutic impact of currently available antioxidants in cardiovascular disease, we developed a nanomedicine-based delivery system for the O2(-)-scavenging enzyme copper/zinc superoxide dismutase (CuZnSOD), in which CuZnSOD protein is electrostatically bound to a poly-l-lysine (PLL50)-polyethylene glycol (PEG) block copolymer to form a CuZnSOD nanozyme. Various formulations of CuZnSOD nanozyme are covalently stabilized by either reducible or nonreducible crosslinked bonds between the PLL50-PEG polymers. Herein, we tested the hypothesis that PLL50-PEG CuZnSOD nanozyme delivers active CuZnSOD protein to neurons and decreases blood pressure in a mouse model of angiotensin II (AngII)-dependent hypertension. As determined by electron paramagnetic resonance spectroscopy, nanozymes retain full SOD enzymatic activity compared to native CuZnSOD protein. Nonreducible CuZnSOD nanozyme delivers active CuZnSOD protein to central neurons in culture (CATH.a neurons) without inducing significant neuronal toxicity. Furthermore, in vivo studies conducted in adult male C57BL/6 mice demonstrate that hypertension established by chronic subcutaneous infusion of AngII is significantly attenuated for up to 7 days after a single intracerebroventricular injection of nonreducible nanozyme. These data indicate the efficacy of nonreducible PLL50-PEG CuZnSOD nanozyme in counteracting excessive O2(-) and decreasing blood pressure in AngII-dependent hypertensive mice after central administration. Additionally, this study supports the further development of PLL50-PEG CuZnSOD nanozyme as an antioxidant-based therapeutic option for hypertension.
Klyachko, N.L., Haney, M.J., Zhao, Y., Manickam, D.S., Mahajan, V., Suresh, P., Hingtgen, S.D., Mosley, R.L., Gendelman, H.E., Kabanov, A.V., Batrakova, E.V.
AIMS: Active targeted transport of the nanoformulated redox enzyme, catalase, in macrophages attenuates oxidative stress and as such increases survival of dopaminergic neurons in animal models of Parkinson’s disease. Optimization of the drug formulation is crucial for the successful delivery in living cells. We demonstrated earlier that packaging of catalase into a polyion complex micelle (‘nanozyme’) with a synthetic polyelectrolyte block copolymer protected the enzyme against degradation in macrophages and improved therapeutic outcomes. We now report the manufacture of nanozymes with superior structure and therapeutic indices.
METHODS: Synthesis, characterization and therapeutic efficacy of optimal cell-based nanoformulations are evaluated.
RESULTS: A formulation design for drug carriers typically works to avoid entrapment in monocytes and macrophages focusing on small-sized nanoparticles with a polyethylene glycol corona (to provide a stealth effect). By contrast, the best nanozymes for delivery in macrophages reported in this study have a relatively large size (≈ 200 nm), which resulted in improved loading capacity and release from macrophages. Furthermore, the cross-linking of nanozymes with the excess of a nonbiodegradable linker ensured their low cytotoxicity, and efficient catalase protection in cell carriers. Finally, the ‘alternatively activated’ macrophage phenotype (M2) utilized in these studies did not promote further inflammation in the brain, resulting in a subtle but statistically significant effect on neuronal regeneration and repair in vivo.
CONCLUSION: The optimized cross-linked nanozyme loaded into macrophages reduced neuroinflammatory responses and increased neuronal survival in mice. Importantly, the approach for nanoformulation design for cell-mediated delivery is different from the common requirements for injectable formulations.
Zhao Y., Alakhova D.Y., Kim J.O., Bronich T.K., Kabanov A.V.
The antitumor efficacy of Doxil® is hindered by the poor release of the active drug from the liposome at the tumor sites. This study investigates a possibility to enhance drug release from the liposomes and increase therapeutic efficacy of Doxil® by administering Pluronic block copolymers once the liposomal drug accumulates in the tumor sites. In our study, the fluorescence de-quenching experiments were designed to investigate the drug release from liposome by Pluronic P85. MTT cytotoxicity assay and confocal microscopy images were carried out to determine whether Pluronic P85 could facilitate release of Dox from Doxil®. Anti-tumor growth and distribution of drug were evaluated when Pluronic P85 was injected 1h, 48h, or 96h after the Doxil® administration in A2780 human ovarian cancer xenografts. Addition of Pluronic P85 resulted in release of Dox from the liposomes accompanied with significant increases of Dox delivery and cytotoxic effect in cancer cells. The greatest anti-tumor effect of single injection of Doxil® was achieved when Pluronic P85 was administered 48h after Doxil®. The confocal tile scanning images of tumor section showed that copolymer treatment induced the release of the drug in the tumors from the vessels regions to the bulk of the tumor. No release of the drug remaining in circulation was observed. Our study has demonstrated a simple approach for localized release of Dox from liposome by Pluronic P85 at the tumor site, which was therapeutically beneficial.
Send us an e-mail and we will get back to you in 24 hours.