Oligomeric Hyaluronic Acid Research Articles

Deuterated colchicine liposomes based on oligomeric hyaluronic acid modification enhance anti-tumor effect and reduce systemic toxicity

Deuterated drugs are produced by substituting hydrogen atoms with deuterium atoms at specific sites in a drug molecule to prolong its metabolic cycle and reduce the production of toxic metabolites. Deuterated drugs have recently attracted increasing attention from the pharmaceutical industry. Colchicine exhibits a strong anti-tumor activity but has a short half-life, rapid attenuated drug concentration, narrow treatment window, and lack of tumor-specific targeting in vivo, resulting in toxicity and side effects. In this study, we explored whether deuteration could reduce the toxicity of colchicine. We prepared deuterated colchicine liposomes coated with oligo-hyaluronic acid, which can bind to the tumor-specific CD44 receptor and reduce the clearance of immune cells from the blood, resulting in a long blood circulation time and active targeting. We observed that deuteration of the colchicine B ring reduced drug toxicity and improved the anti-tumor response in 4 T1 breast cancer. Liposomes modified with oligo-hyaluronic acid exhibited increased tumor accumulation, further improving the anti-tumor effect of the drugs. Our results provide a basis for the development and application of deuterated drugs in the field of nano-preparations.

Abstract 5399: Synergistic combination nanomedicine of doxorubicin and oligo hyaluronic acid inhibits DNA damage repair and overcomes drug resistance in metastatic triple-negative breast cancer

Abstract Background: Triple-negative breast cancer (TNBC) is a highly aggressive breast cancer subtype. Despite the early response to chemotherapy, high incidence of recurrence leads to short overall survival and poor prognosis. Native hyaluronic acid (HA) interacts with CD44 and receptors for hyaluronan mediated motility (RHAMM) overexpressed in TNBC mediating chemo-resistance and metastasis. In contrast, oligomeric HA (oHA) can disrupt HA-CD44/RHAMM interactions and attenuate oncogenic and pro-metastatic pathways, such as the mitogen-activated protein kinase/extracellular-signal-regulated kinase (MAPK/ERK) pathway, reducing downstream DNA repair and drug resistance markers. Our previous work demonstrated that oHA can exert synergistic anti-tumor and anti-metastatic effects when combined with doxorubicin (DOX) and co-loaded in an integrin-targeted iRGD-modified nanoparticle system (iRGD-DOX-oHA-PLNs). This study aims to investigate the inhibitory effect of iRGD-DOX-oHA-PLNs on both DNA single-strand break (SSB) and double-strand break (DSB) repair proteins and drug efflux pumps responsible for multidrug resistance to enhance DOX efficacy in breast cancer gene 1 (BRCA1) mutant and non-mutant TNBC. Methods: The cytotoxicity of DOX, oHA and their combinations in free solution or in nanoparticles was evaluated by clonogenic assay in human MDA-MB-231-luc-D3H2LN and MDA-MB-436 (BRCA1 mutant) TNBC cells. The in vitro expression of a DNA DSB marker, DNA repair markers, and drug efflux pump P-glycoprotein (P-gp) were measured by Western blot. The in vivo expression level of BRCA1 and Rad51 in an orthotopic TNBC mouse model was determined by immunohistochemical staining. Results: The combination of DOX-oHA showed synergism against both BRCA1 mutant and non-mutant TNBC cells. The iRGD-DOX-oHA-PLNs induced great increases in DNA DSBs demonstrated by the highest γH2AX level compared to other treatment groups. These nanoparticles also showed inhibitory effects on the expression of both DNA SSB repair protein (poly (ADP-ribose) polymerase) and DNA DSB repair proteins (Rad50 and Rad51), contributing to the enhanced efficacy of chemotherapy. The immunohistochemical staining of tumor tissues indicated lower levels of BRCA1 and Rad51 after iRGD-DOX-oHA-PLN treatment than the formulation without oHA, attributable to the effect of intracellularly delivered oHA on limiting the MAPK signaling. Additionally, iRGD-DOX-oHA-PLNs reduced the expression of the drug efflux pump P-gp as compared to DOX treatment groups without oHA. Conclusion: The co-delivery of oHA and DOX in the iRGD-DOX-oHA-PLNs efficiently blocked DNA damage repair and down-regulated the drug efflux pump P-gp, thus improving the efficacy of DOX. Collectively, this nanoparticle system could be a promising option for metastatic TNBC treatment. Citation Format: Pei Zhi, Ibrahim Alradwan, Tian Zhang, HoYin Lip, Abdulmutalib Zetrini, Chunsheng He, Jeffery Henderson, Andrew Michael Rauth, Xiao Yu Wu. Synergistic combination nanomedicine of doxorubicin and oligo hyaluronic acid inhibits DNA damage repair and overcomes drug resistance in metastatic triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5399.

A comparison study of hyaluronic acid hydrogel exquisite micropatterns with photolithography and light-cured inkjet printing methods

Abstract The microstructure design of hydrogel materials offers a broad range of practical applications and is extensively used in flexible sensors, polymer microneedles, microfluidic chips, and other biomedical engineering fields. Among the bio-sourced hydrogels, oligomeric hyaluronic acid (HA) possesses wound healing, anti-tumor, and angiogenesis properties. However, micropatterning soft hydrogels, such as HA-relative hydrogels containing 90% water by weight, continue to pose difficulties for both high precision and micro-scale lithography. The purpose of this study was to compare the photolithography and light-cured inkjet printing methods of methacryloyl HA hydrogel (HAMA-gel) to those for synthetic light-curable polymer resins. Photolithography and light-cured inkjet printing methods with designed scale, high resolution, and little processing times were used to effectively prepare micropatterns of HAMA-gel. The well-shaped micropatterns consisted of parallel channels in tens of micrometers and strip/grid lines in the hundreds of micrometers. Human vein endothelial cells cultured on the material’s surface demonstrated that HAMA-gel had good biocompatibility. The width of the flow channel (10 and 20 µm) was regulated on the surface of the microstructure to allow for simultaneous control of cell growth along the flow channel and groove directions.

Targeting Signaling Pathways of Hyaluronic Acid and Integrin Receptors by Synergistic Combination Nanocomposites Inhibits Systemic Metastases and Primary Triple Negative Breast Cancer

AbstractTriple negative breast cancer (TNBC) has a poor prognosis due to its aggressive nature, high incidence of distant metastasis, and lack of targets for effective therapy. Therefore, a novel multifunctional biopolymer‐anticancer drug combination nanomedicine is designed for the prevention of spontaneous metastasis while treating primary TNBC. Oligomeric hyaluronic acid (oHA) and doxorubicin (DOX) at a synergistic ratio against TNBC cells are co‐loaded in a polymer‐lipid hybrid nanoparticle (PLN) which is then functionalized with an internalizing cyclic peptide iRGD (iRGD‐DOX‐oHA‐PLN). iRGD conjugation enhances cellular uptake and cytotoxicity in vitro and nanoparticle (NP) accumulation in human TNBC tumors that overexpress integrins. NP‐delivered oHA inhibits cell migration and invasion in vitro via the intracellular release of oHA that interacts with the receptor for hyaluronan mediated motility and downregulates the phospho‐extracellular signal‐regulated kinase (p‐ERK) signaling pathway. Intravenously injected iRGD‐DOX‐oHA‐PLN significantly inhibits the growth of primary TNBC tumors in a mouse model and prevents spontaneous metastasis to the lungs and lymph nodes, superior to free solutions of DOX, oHA, or both and NP formulations loaded with DOX or oHA. These results suggest that iRGD‐DOX‐oHA‐PLN can be a promising bioactive polymer‐drug combination nanomedicine for the treatment of TNBC and prevention of its spontaneous metastasis.

An electrochemiluminescence sensor of magnetic glycosyl-imprinted microspheres based on multi-probe signal amplification for CD44v6 determination

A CD44v6 detection method was developed by using glycosyl-imprinted technique and enhanced with multi-probe signal amplification. Oligomeric hyaluronic acid was used as the template molecule, which was prepared by the cleavage of polyhyaluronic acid using hyaluronidase. Mercaptophenylboronic acid was self-assembled on the surface of core-shell [email protected]3O4 magnetic microspheres through the covalent interaction between its thio group and Au. Then oligomeric hyaluronic acid (<10 kDa) was linked on [email protected]3O4 via the formation of a cyclic ester between boronic acid and the ortho-dihydroxy group of oligomeric hyaluronic acid. By using oligomeric hyaluronic acid as a template and p-aminophenylboronic acid (p-APBA) as the functional monomer, boronate affinity glycosyl-imprinted polymer films were prepared on [email protected]3O4 by chemical polymerization. The magnetic glycosyl-imprinted polymer microspheres (MGIP) were attached on the surface of the magnetron-controlled glassy carbon electrode (MGCE) as the recognition component rebinding for the template, and CD44v6 was specifically recognized by the binding of the imprinted cavities to the oligomeric hyaluronic acid. The boronic acid functionalized tris-(2,2’-bipyridyl) ruthenium (II) (Ru(bpy)3-BO3) was used as a molecular probe to label the adsorbed oligomeric hyaluronic acid of CD44v6. With the aid of the multiply captured Ru(bpy)32+ probes on CD44v6 and the high luminescence efficiency of Ru(bpy)32+, the electrochemiluminescence response of the sensor exhibits a high sensitivity for CD44v6 determination with a detection limit of 0.08 ng/L. The method was applied in the assay of CD44v6 in serum samples.

A paclitaxel and microRNA-124 coloaded stepped cleavable nanosystem against triple negative breast cancer

BackgroundTriple negative breast cancer (TNBC) is one of the most biologically aggressive breast cancers and lacks effective treatment options, resulting in a poor prognosis. Therefore, studies aiming to explore new therapeutic strategies for advanced TNBC are urgently needed. According to recent studies, microRNA-124 (miR124) not only inhibits tumour growth but also increases the sensitivity of TNBC to paclitaxel (PTX), suggesting that a platform combining PTX and miR124 may be an advanced solution for TNBC.ResultsHerein, we constructed a stepped cleavable calcium phosphate composite lipid nanosystem (CaP/LNS) to codeliver PTX and miR124 (PTX/miR124-NP). PTX/miR124-NP exhibited superior tumor microenvironment responsive ability, in which the surface PEG layer was shed in the mildly acidic environment of tumor tissues and exposed oligomeric hyaluronic acid (o-HA) facilitated the cellular uptake of CaP/LNS by targeting the CD44 receptor on the surface of tumor cells. Inside tumour cells, o-HA detached from CaP/LNS due to the reduction of disulfide bonds by glutathione (GSH) and inhibited tumour metastasis. Then, PTX and miR124 were sequentially released from CaP/LNS and exerted synergistic antitumour effects by reversing the Epithelial-Mesenchymal Transition (EMT) process in MDA-MB-231 cells. Moreover, PTX/miR124-NP showed significant antitumour efficiency and excellent safety in mice bearing MDA-MB-231 tumours.ConclusionBased on these results, the codelivery of PTX and miR124 by the CaP/LNS nanosystem might be a promising therapeutic strategy for TNBC.

Reversing P-Glycoprotein-Associated Multidrug Resistance of Breast Cancer by Targeted Acid-Cleavable Polysaccharide Nanoparticles with Lapatinib Sensitization.

For reversing the treatment failure in P-glycoprotein (P-gp)-associated MDR (multidrug resistance) of breast cancer, a high dose of Lapatinib (Lap), a substrate of breast cancer-resistant protein, was encapsulated into safe and effective acid-cleavable polysaccharide-doxorubicin (Dox) conjugates to form targeted HPP-Dox/Lap nanoparticles with an optimal drug ratio and appropriate nanosize decorated with oligomeric hyaluronic acid (HA) for specially targeting overexpressed CD44 receptors of MCF-7/ADR. The markedly increased cellular uptake and the strongest synergetic cytotoxicity revealed the enhanced reversal efficiency of HPP-Dox/Lap nanoparticles with reversal multiples at 29.83. This was also verified by the enhanced penetrating capacity in multicellular tumor spheroids. The reinforced Dox retention and substantial down-regulation of P-gp expression implied the possible mechanism of MDR reversal. Furthermore, the efficient ex vivo accumulation and distribution of nanoparticles in the tumor site and the high tumor growth inhibition (93%) even at a lower dosage (1 mg/kg) as well as lung metastasis inhibition in vivo with negligible side effects revealed the overwhelming advantages of targeted polysaccharide nanoparticles and Lap-sensitizing effect against drug-resistant tumor. The development of an efficient and nontoxic-targeted polysaccharide delivery system for reversing MDR by synergistic therapy might provide a potential clinical application value.

Novel dual ROS-sensitive and CD44 receptor targeting nanomicelles based on oligomeric hyaluronic acid for the efficient therapy of atherosclerosis

Although the clinical usage of drugs administration was raising, the application of nanoparticles encapsulating the hydrophobic drugs with plummy efficiency was very scarce for atherosclerosis (AS) treatment. In this work, a novel dual ROS-sensitive and CD44 receptors targeting amphiphilic carrier material, oligomeric hyaluronic acid-2′-[propane-2,2-diyllbls (thio)] diacetic acl-hydroxymethylferrocene (oHA-TKL-Fc), named HASF, was synthesized and characterized by 1H-NMR spectra. Then, we combined curcumin (Cur) with HASF into nano-micelles (HASF@Cur micelles) by self-assembling method. The resulting HASF@Cur micelles had the average size of 150.8 nm and zeta potential of -35.04 mV to maintain the will-defined spheroidal structure and stability. Importantly, the HASF@Cur micelles had ultrahigh entrapment efficiency (about 51.41 %). Moreover, in vitro release study, Cur release from HASF@Cur micelles was effective in the reactive oxygen species (ROS) condition, and the release rate was interrelated with the concentration of hydrogen peroxide (H2O2). Further, fluorescence imaging showed that the HASF@Cur micelles could more selective access to Raw 264.7 cells than free Cur via oHA-receptor mediated endocytosis. The MTT assay attested the safety of amphiphilic carrier material HASF. Additionally, the results of in vivo Oil red O lipid staining studies showed that the lesion area of the aorta was reduced to 47.3±3.4 % with HASF@Cur micelles, compared with the lesion area of Cur group (63.2±2.7 %), HASF@Cur micelles had the more remarkable effect in reducing lesion area (*P < 0.05). Consequently, the novel dual ROS-sensitive and CD44 receptors targeting drug delivery system would become a promising strategy for atherosclerosis.

Development of dual-targeted nano-dandelion based on an oligomeric hyaluronic acid polymer targeting tumor-associated macrophages for combination therapy of non-small cell lung cancer

In this study, the novel carrier materials were screened to structure targeting nano-micelles (named ‘nano-dandelion’) for synchronous delivery of curcumin (Cur) and baicalin (Bai), which could effectively overcome the tumor resistance. Mannose (Man) was found to bind better to CD206 receptors on the surface of tumor-associated macrophages (TAMs), thereby increasing the number of nano-dandelion engulfed by TAMs. Furthermore, oligomeric hyaluronic acid (oHA) was able to target CD44 receptors, resulting in recruitment of a higher number of nano-dandelion to locate and engulf tumor cells. The disulfide bond (S–S) in 3,3′-dithiodipropionic acid (DA) could be broken by the high concentration of glutathione (GSH) in the tumor microenvironment (TME). Based on this, we selected DA to connect hydrophobic fragments (quercetin, Que) and oHA. A reduction-sensitive amphiphilic carrier material, quercetin–dithiodipropionic acid–oligomeric hyaluronic acid–mannose–ferulic acid (Que–S–S–oHA–Man–FA; QHMF) was fabricated and synthesized by 1H NMR. Next, QHMF self-assembled into nano-dandelion, i.e. encapsulated Cur and Bai in water. Critical experimental conditions in the preparation process of nano-dandelion that could affect its final properties were explored. Nano-dandelion with a small particle size (121.0 ± 15 nm) and good normal distribution (PI = 0.129) could easily enter tumor tissue through vascular barrier. In addition, nano-dandelion with a suitable surface potential (–20.33 ± 4.02 mV) could remain stable for a long duration. Furthermore, good cellular penetration and tumor cytotoxicity of nano-dandelion were demonstrated through in vitro cellular studies. Finally, effective antitumor activity and reduced side effects were confirmed through in vivo antitumor experiments in A549 tumor-bearing nude mice.

Enhanced In Vitro and In Vivo Anticancer Properties by Using a Nanocarrier for Co-Delivery of Antitumor Polypeptide and Curcumin.

In this paper, a novel pH and redox dual-sensitive nanocarrier loaded with curcumin (Cur) and anticancer polypeptide (AP) was developed for dual targeting mitochondrial and CD44 receptor. The amphiphilic block copolymer was prepared by triphenylphosphonium (TPP)/oligomeric hyaluronic acid (oHA)/disulfide-menthone 1,2-glycerol ketal (SM), hereinafter referred to as TPP-oHSM. The TPP targeted the mitochondria, pH/redox dual-sensitive SM served as a hydrophobic part, and the CD44 receptor targeting oHA worked as a hydrophilic part. The chemical structure of the TPP-oHSM was identified using 1H NMR and FTIR technologies. Cur and AP were loaded into the TPP-oHSM micelles by self-assembly and denoted as C/A@TM. The C/A@TM prepared in this study exhibited an approximately spherical structure, with a mean diameter of 191.3 ± 3.1 nm and a negative zeta potential of -26.10 ± 0.45 mV. The in vitro release study and cellular uptake test revealed that the C/A@TM targeted the mitochondria and CD44 receptor, as well as it showed sensitivity towards pH and redox. In addition, the C/A@TM demonstrated satisfactory cytotoxic effects against MDA-MB-231 cells and MCF-7 cells. Finally, the in vivo application of the C/A@TM showed excellent therapeutic effects. The C/A@TM developed in this study exhibited promising multifunctional properties as a co-delivery carrier of polypeptide and chemical drug for an effective clinical therapy for cancer.

LAPONITE® nanoplatform functionalized with histidine modified oligomeric hyaluronic acid as an effective vehicle for the anticancer drug methotrexate.

The synthetic clay material, LAPONITE® (LAP), having a nanodisk structure together with a negatively charged surface, has been used for effective drug encapsulation by virtue of its interlayer space. In this research effort, the LAP nanodisk was used for the first time to encapsulate the antifolic methotrexate (MTX); the MTX-loaded LAP nanodisks (LAP/MTX) demonstrated a high drug loading efficiency of 80.39%. An efficient and reliable tumor-targeting device that rests on the synthesized oligomeric hyaluronic acid-l-histidine (oHA-His) was then encapsulated in the MTX-loaded LAP disks (forming LAP/MTX/oHA-His nanohybrids). The drug released from the LAP/MTX/oHA-His nanohybrids was pH-dependent and matched the first-order kinetics that describes the diffusion mechanism. In vitro biological evaluation manifested that the MTX-loaded LAP nanocarriers, particularly the LAP/MTX/oHA-His nanohybrids that have targetability and lysosomal antineoplastic activity, can be effectively internalized by the MCF-7 cell line, and can exhibit a more prominent anticancer cytotoxicity than free MTX. In vivo studies with mice indicated that the LAP/MTX/oHA-His nanohybrids demonstrated much higher antitumor efficiency compared to the LAP/MTX nanohybrids and pure MTX. Taken together, the LAP/oHA-His, CD44 receptor targeting and pH-sensitive multifunctional nanohybrids conferred the MTX with excellent cytocompatibility, dispersion stability, sustained pH-responsive release properties, and improved anticancer activity, and may be further developed as a potential active nanoplatform for various anticancer drugs.

Coadministration of Oligomeric Hyaluronic Acid-Modified Liposomes with Tumor-Penetrating Peptide-iRGD Enhances the Antitumor Efficacy of Doxorubicin against Melanoma.

A safe and efficient tumor-targeting strategy based on oligomeric hyaluronic acid (HA) modification and coadministration of tumor-penetrating peptide-iRGD was successfully developed. In this study, common liposomes (cLip) were modified by oligomeric HA to obtain HA-Lip. After injection into rats, HA-Lip showed good stealth in the bloodstream and lower liver distribution compared with cLip. Moreover, our HA-Lip could be internalized into B16F10 cells (CD44-overexpressing tumor cells) through HA-CD44 interaction. After systemic administration to B16F10 melanoma-bearing mice, HA-Lip showed an increased distribution in tumor due to the prolonged blood circulation time and the enhanced penetration and retention effect. When coadministered with iRGD, the tumor penetration of HA-Lip was significantly enhanced, which could promote HA-Lip internalization by tumors cells located in deep tumor regions through receptor-mediated endocytosis. Furthermore, doxorubicin (DOX)-loaded HA-Lip coadministering with iRGD showed much better antitumor effect compared to DOX-loaded cLip and DOX-loaded cLip in combination with iRGD. In systemic toxicity test, DOX-loaded HA-Lip could significantly decrease the cardiotoxicity and myelosuppression of DOX. Taken together, our results demonstrated that coadministration of oligomeric HA-modified liposomes with iRGD could be a promising treatment strategy for targeted therapy of melanoma.