Benzylamide Groups Research Articles

Theoretical investigations on copper catalyzed C N cross-coupling reaction between aryl chlorides and amines

Abstract The mechanism of CuI/oxalic diamide catalyzed arylation of amines with chlorobenzene was investigated computationally. CuI/N,N′-bis(2,4,6-trimethoxyphenyl)-oxalamide (BTMPO) system was selected as model catalytic system. On the basis of relative Gibbs free energy, five different possible intermediate Cu1 complexes were selected for mechanistic studies. Intermediate complexes Int1–Int3 have a BTMPO ligand deprotonated at one or both NH groups and a benzylamine or benzylamide group coordinated to Cu1. Intermediate Int4 has a mono deprotonated BTMPO and an ƞ2-coordinated chlorobenzene ligand. Int5 is anionic analogue of Int4 in which both NH groups of the BTMPO ligand are deprotonated. DFT calculations were carried out for oxidative addition/reductive elimination and single electron transfer mechanisms. Int2 shows the lowest activation energy barrier (23.5 kcal/mol) for oxidative addition/reductive elimination mechanism. All the intermediates show higher activation energy for single electron transfer mechanism than oxidative addition/reductive elimination mechanism of Int2.

Design, synthesis, and biological evaluation of novel 1-oxo-1,2,3,4-tetrahydropyrazino[1,2-a]indole-3-carboxamide analogs in MCF-7 and MDA-MB-468 breast cancer cell lines

A series of novel 1-oxo-1,2,3,4-tetrahydropyrazino[1,2-a]indole-3-carboxamide analogs were designed and synthesized for developing pyrazinoindolone scaffolds as anti-breast cancer agents. Compounds 1h and 1i, having a furan-2-yl-methylamide and benzylamide group, respectively, exhibited more potent cytotoxicity in MDA-MB-468 triple-negative breast cancer (TNBC) cells than compounds possessing aliphatic groups. Compounds 2a and 2b, as (R)-enantiomers of 1h and 1i, also had inhibitory activity against MDA-MB-468 cells. Moreover, analogs (3a–b and 3d–e) bearing a benzyl group at the N-2 position showed more potent activity than gefitinib, as a potent EFGR-TK inhibitor. Especially, compound 3a exhibited selective cytotoxic activity against MDA-MB-468 cells; it also had a synergistic effect in combination with gefitinib against MDA-MB-468 cells. In addition, we confirmed that compounds 3a and 3d inhibit phosphorylation of Akt in MDA-MB-468 cells using western blot analysis. Therefore, these 1-oxo-1,2,3,4-tetrahydropyrazino[1,2-a]indole-3-carboxamide analogs may be helpful for investigating new anti-TNBC agents.

Enhancement of the biological activity of BMP‐2 by synthetic dextran derivatives

In the present study, we explored the binding capacity of synthetic heparin-like dextran derivatives to recombinant human bone morphogenetic protein 2 (BMP-2), a heparin-binding osteoinductive growth factor. Affinity electrophoresis analysis provided evidence that carboxymethylated dextran polymers grafted with high amounts of benzylamide groups (named DMCB) interact with BMP-2. The capability of such polysaccharides to potentiate the growth factor biological activity was then investigated. In vitro, DMCB dose-dependently promoted osteoblast differentiation induced by BMP-2 in C2C12 myoblasts more efficiently than heparin. A screening study provided evidence that the potentiating effects of the dextran derivatives on the BMP-2-induced alkaline phosphatase activity improved with their benzylamide groups content and, therefore, with their affinity for the growth factor. The biological activity of BMP-2 was monitored in the culture medium after 6 days using C2C12 cells (containing a BMP sensitive luciferase reporter gene). Like heparin, DMCB sustained the biological activity of the growth factor; this result suggests that the formation of the BMP-2/DMCB complex may protect the protein from being inactivated. In rats in vivo, DMCB also stimulated ectopic calcification mediated by BMP-2. These data indicate that dextran-based polysaccharides prolong the half-life of the growth factor and promote its biological activity.

“Fingertip”-Guided Noncovalent Functionalization of Carbon Nanotubes by Dendrons

Noncovalent functionalization of carbon nanotubes (CNTs) by dendrons was demonstrated. Certain types of dendrons successfully functionalized CNT surfaces through the noncovalent interactions between the peripheries of the dendrons and the sidewalls of CNTs. Dendrons have a unique anisotropic shape and an orthogonal functional group at their apex, and thus can generate a certain spacing between the functional groups upon immobilization on surfaces. Atomic force microscope (AFM) imaging, dispersion experiments, and MicroRaman spectroscopy were employed for the characterization of the functionalization. The binding was found to be governed by the chemical nature of the terminal groups, namely, the "fingertips", through a comparison study on the adsorption efficiency of the dendron analogs. Functional groups such as the carboxylic acid group and the benzyl amide group were effective for the cooperative binding. AFM analysis showed that the average spacing generated by the dendrons was 14-15 nm at a particular adsorption condition. Assembling streptavidin on the tubes through the dendrons and biotin confirmed the realization of the regulated spacing as well as the elimination of unwanted aggregation. The noncovalent functionalization of CNTs by a dendron can be a new approach toward sensible nanobiodevices, not only by introducing biomolecular probes on CNTs without disruption of the electronic network of the tubes, but also by providing the immobilized probe molecules with a space ample enough to minimize steric hindrance for the unhindered interaction with their target species.

Dextran derivatives modulate collagen matrix organization in dermal equivalent

Dextran derivatives can protect heparin binding growth factor implied in wound healing, such as transforming growth factor-β1 (TGF-β1) and fibroblast growth factor-2 (FGF-2). The first aim of this study was to investigate the effect of these compounds on human dermal fibroblasts in culture with or without TGF-β1. Several dextran derivatives obtained by substitution of methylcarboxylate (MC), benzylamide (B) and sulphate (Su) groups were used to determine the effects of each compound on fibroblast growth in vitro. The data indicate that sulphate groups are essential to act on the fibroblast proliferation. The dextran derivative LS21 DMCBSu has been chosen to investigate its effect on dermal wound healing process. Fibroblasts cultured in collagenous matrices named dermal equivalent were treated with the bioactive polymer alone or associated to TGF-β1 or FGF-2. Cross-sections of dermal equivalent observed by histology or immunohistochemistry, demonstrated that the bioactive polymer accelerates the collagen matrices organization and stimulates the human type-III collagen expression. This bioactive polymer induces apoptosis of myofibroblast, property which may be beneficial in treatment of hypertrophic scar. Culture media analyzed by zymography and Western blot showed that this polymer significantly increases the secretion of zymogen and active form of matrix metalloproteinase-2 (MMP-2), involved in granulation tissue formation. These data suggest that this bioactive polymer has properties which may be beneficial in the treatment of wound healing.

Growth inhibition of MCF-7 tumor cell line by phenylacetate linked to functionalized dextran

We investigated the antiproliferative effect of phenylacetate covalently linked to dextran derivatives (DMCBPA conjugates) on human breast cancer MCF-7 cells. We show that free sodium phenylacetate (NaPA) inhibits the cell growth (IC50 = 14 mM), while an important inhibitory effect is observed for DMCBPA conjugates. The IC50 dose of these conjugates is as low as 1.0 mg/ml, corresponding to 1.3 mM of phenylacetate. The precursors, dextran substituted with methylcarboxylate and benzylamide groups, did not affect the growth of MCF-7 tumor cells. We have observed that MCF-7 cell growth inhibition depends on amount of phenylacetate linked to the conjugate. The data indicated that an optimum antiproliferative effect is more significant when the amount of phenylacetate groups present on the dextran backbone is high. Analysis of doubling time by growth kinetics study shows that conjugates have more time-sustained effect than free NaPA. It is noteworthy that the inhibitory effect is observed at non-toxic concentration. Theses conjugates could be considered as acceptable derivatives to prevent tumor progression.

Effect of a dextran derivative associated with TGF-β1 or FGF-2 on dermal fibroblast behaviour in dermal equivalents

Dextran derivatives that mimic the action of heparin have been shown to protect heparinbinding growth factors, such as Transforming Growth Factor-β1 (TGF-β1) and Fibroblast Growth Factor-2 (FGF-2). The aim of this study was to investigate the effect of LS21 DMCBSu, a dextran derivative which contains methylcarboxylate, benzylamide and sulfate groups, both by itself and when combined with TGF-β1 and FGF-2, on the behaviour of fibroblasts. Two systems were assessed: a monolayer culture and three-dimensional collagenous matrices (dermal equivalent). Polymeric biomaterial LS21 DMCBSu and LS21 DMCBSu associated with either TGF-β1 or FGF-2, were added to the monolayer culture on day 3. After 7 days of culture the number of cells was determined. Two treatments were carried out on the dermal equivalents: 9 days of treatment from day 0 to day 9 of culture and 9 days of treatment from day 21 to day 30 of culture for the premature and the mature dermal equivalents respectively. In the monolayer culture, the bioactive polymer produced a slight increase in fibroblast growth (10% with 10 μg/ml of LS21 DMCBSu) and promoted the stimulating effect of the growth factors on cell growth. In the premature dermal equivalents growth was stimulated by 20% when 10 μg/ml LS21 DMCBSu was added. The dextran derivative mixed with TGF-β1slightly inhibited the growth effect of the growth factor in the dermal equivalents. The functionalized dextran with FGF-2 enhanced the stimulating effect of the growth factor in the premature dermal equivalent. A significant increase in cell growth was observed with the fibroblasts treated with the FGF-2 LS21 DMCBSu mixture and FGF-2 (51% and 40%, respectively). However, none of the described treatments affected the cell growth in the mature dermal equivalent. Furthermore, the dextran derivative had no effect on dermal contraction under these experimental conditions (3D culture).

Heparin-like dextran derivatives as well as glycosaminoglycans inhibit the enzymatic activity of human cathepsin G

Some synthetic dextran derivatives that mimic the action of heparin/heparan sulfate were previously shown to inhibit neutrophil elastase and plasmin. Here we report that these derivatized dextrans also inhibit cathepsin G (CatG). Dextran containing carboxymethyl and benzylamide groups (RG1150) as well as those containing carboxymethyl, sulfate and benzylamide groups (RG1192), were the most efficient inhibitors of CatG activity. RG1192 and RG1150 bind CatG with a Ki of 0.11 and 0.17 nM, respectively, while carboxymethylated sulfated dextran (RG1503) as well as heparin, heparan sulfate and dermatan sulfate bind CatG with a 7- to 30-fold lower affinity. Variation of Ki with ionic strength indicates that ionic interactions account for 26% of the RG1503–CatG binding energy, while binding of RG1192 or RG1150 to CatG is mainly governed by non-electrostatic interactions. This, together with the fact that these compounds both protect fibronectin and laminin against CatG-mediated degradation, suggest that specific dextran derivatives can contribute to the regulation of CatG activity.

Poly( N-ethyl-4-vinylpyridinium) bromide as a potential probe to select heparin-like anticoagulant polyanions

Carboxymethyl dextran benzylamide sulfates compounds are polyanionic synthetic dextran derivatives with a random distribution of glucosyl units substituted with carboxymethyl, benzylamide, and sulfate groups. Two samples of derivatized dextrans with similar carboxymethyl and benzylamide contents, and with different sulfate content and one sample without benzylamide groups were prepared. The anticoagulant activity of the samples was estimated by the capacity to inhibit procoagulant activity of thrombin, following which the inhibition of this activity was monitored by addition of the polycation poly( N-ethyl-4-vinylpyridinium) bromide (PEVP). The potent inhibition by PEVP strongly suggests that the anticoagulant activity of the derivatives is attributable to a small portion of their overall negative charges. The comparison study of the samples implies that specific sites capable to bind thrombin are composed of rather small number of sulfate groups recognized and blocked by PEVP. These findings demonstrate high potential of PEVP as a probe for screening of heparin-like anticoagulant polyanions.

Anticoagulant properties of dextranmethylcarboxylate benzylamide sulfate (DMCBSu); a new generation of bioactive functionalized dextran

Dextranmethylcarboxylate benzylamide sulfate (DMCBSu), a functionalized dextran, exhibits anticoagulant properties. Its synthesis involves three steps: a carboxymethylation with monochloroacetic acid in alkaline water–isopropanol, a benzylamidification of some of the methylcarboxylate groups with benzylamine in the presence of a water soluble carbodiimide and a partial sulfation of the remaining hydroxyl groups with SO 3–pyridine in dimethylformamide. This procedure yields reproducibly DMCBSu with degrees of substitution in methylcarboxylate (MC), benzylamide (B) and sulfate (Su) groups, respectively, up to 1.61, 0.35 and 1.5, each obtained in one step. For a degree of substitution of methylcarboxylate ca. 1, the presence of sulfate groups is absolutely necessary to confer anticoagulant activities to the samples. In addition, the anticoagulant ability is higher for derivatives bearing benzylamide groups. The anticoagulant ability of DMCBSu increases with the degree of sulfation, reaching 20% of heparin activity for a degree of substitution of Su groups about 1.3.

Human Plasmin Enzymatic Activity Is Inhibited by Chemically Modified Dextrans

Some synthetic dextran derivatives that mimic the action of heparin/heparan sulfate were shown to promote in vivo tissue repair when added alone to wounds. These biofunctional mimetics were therefore designated as "regenerating agents" in regard to their in vivo properties. In vitro, these biopolymers were able to protect various heparin-binding growth factors against proteolytic degradation as well as to inhibit the enzymatic activity of neutrophil elastase. In the present work, different dextran derivatives were tested for their capacity to inhibit the enzymatic activity of human plasmin. We show that dextran containing carboxymethyl, sulfate as well as benzylamide groups (RG1192 compound), was the most efficient inhibitor of plasmin amidolytic activity. The inhibition of plasmin by RG1192 can be classified as tight binding hyperbolic noncompetitive. One molecule of RG1192 bound 20 molecules of plasmin with a K(i) of 2.8 x 10(-8) m. Analysis with an optical biosensor confirmed the high affinity of RG1192 for plasmin and revealed that this polymer equally binds plasminogen with a similar affinity (K(d) = 3 x 10(-8) m). Competitive experiments carried out with 6-aminohexanoic acid and kringle proteolytic fragments identified the lysine-binding site domains of plasmin as the RG1192 binding sites. In addition, RG1192 blocked the generation of plasmin from Glu-plasminogen and inhibited the plasmin-mediated proteolysis of fibronectin and laminin. Data from the present in vitro investigation thus indicated that specific dextran derivatives can contribute to the regulation of plasmin activity by impeding the plasmin generation, as a result of their binding to plasminogen and also by directly affecting the catalytic activity of the enzyme.

Design of a Gag pentapeptide analogue that binds human cyclophilin A more efficiently than the entire capsid protein: new insights for the development of novel anti-HIV-1 drugs.

Cyclophilin A (hCyp-18), a ubiquitous cytoplasmic peptidyl-prolyl cis/trans isomerase (PPIase), orchestrates HIV-1 core packaging. hCyp-18, incorporated into the virion, enables core uncoating and RNA release and consequently plays a critical role in the viral replication process. hCyp-18 specifically interacts with a single exposed loop of the Gag polyprotein capsid domain via a network of nine hydrogen bonds which mainly implicates a 7-mer fragment of the loop. As previously reported, the corresponding linear heptapeptide Ac-Val-His-Ala-Gly-Pro-Ile-Ala-NH(2) (2) binds to hCyp-18 with a low affinity (IC(50) = 850 +/- 220 microM) but a potentially useful selectivity for hCyp-18 relative to hFKBP-12, another abundant PPIase. On the basis of X-ray structures of Gag fragments:hCyp-18 complexes, we generated a series of modified peptides in order to probe the determinants of the interaction and hence to select a peptidic ligand displaying a higher affinity than the capsid domain of Gag. We synthesized a series of heptapeptides to test the energetic contribution of amino acids besides the Gly-Pro moiety. In particular the importance of the histidine residue for the interaction was underscored. We also investigated the influence of N- and C-terminal modifications. Hexapeptides containing either deaminovaline (Dav) in place of the N-terminal valine or substitution of the C-terminal alanine amide with a benzylamide group displayed increased affinities. Combination of both modifications gave the most potent competitor Dav-His-Ala-Gly-Pro-Ile-NHBn (28) which has a higher affinity for hCyp-18 (K(d) = 3 +/- 0.5 microM) than the entire capsid protein (K(d) = 16 +/- 4 microM) and a very low affinity for hFKBP-12. Some of our results strongly suggest that the title compound is not a substrate of hCyp-18 and interacts preferentially in the trans conformation.

Low-molecular-weight dextran derivatives ( f-CMDB) enter the nucleus and are better cell-growth inhibitors compared with parent CMDB polymers

Carboxymethyldextrans–benzylamide (CMDB) are dextran derivatives that are statistically substituted with carboxymethyl and benzylamide groups. These molecules display a variety of biological effects, one of which is their inhibitory activity against mammary tumor cell growth, both in vitro and in vivo. We and others have previously shown that the effects of CMDB on cell growth are related to their ability to interact with the growth factor FGF-2. The binding modifies the conformation of FGF-2, leading to the suppression of its mitogenic activity. Here, the method previously reported to fragment natural polysaccharide fucans has been applied to CMDB (80,000 g/mol). f-CMDB (fragmented CMDB) of molecular weights from 6000 to 20,000 g/mol were found to be more potent inhibitors of MCF7 mammary tumor cell growth than high-molecular-weight CMDB. Confocal microscopy experiments using CMDB and f-CMDB labeled with the fluorophore DTAF (5-([4,6-dichlorotriazine-2-yl]amino) fluorescein) indicate that only low-molecular-weight f-CMDB penetrate into the nucleus of MCF7 cells. It is thus assumed that the better inhibitory properties demonstrated by f-CMDB, compared with CMDB, are related to their better ability to penetrate the nucleus and interact with nuclear targets, including topoisomerase II. The DNA relaxation properties of the latter are inhibited in vitro by both CMDB and f-CMDB. These findings could help us to develop models of low-molecular-weight oligosaccharide derivatives exhibiting better antiproliferative and antitumor properties.

Effects of the binding of a dextran derivative on fibroblast growth factor 2: secondary structure and receptor-binding studies

CMDB (carboxymethyldextran-benzylamide) are dextrans statistically substituted with carboxymethyl and benzylamide groups which can mimick some of the biological properties of heparin. It has previously been shown that CMDB inhibit autocrine growth of breast tumor cells (Bagheri-Yarmand et al., Biochem Biophys Res Commun 239: 424–428, 1997) and selectively displace fibroblast growth factor 2 (FGF-2) from its receptor. Here, we used circular dichroism and fluorescence anisotropy measurements to show that the conformation of FGF-2 was significantly altered upon its binding to CMDB and to short CMDB fragments prepared within this study. CMDB and fragments formed a stable 1:1 complex with FGF-2, with affinities being estimated as 20 ± 10 nM from fluorescence anisotropy analysis. No such a complex was formed with insulin-like growth factor (IGF-1) or epidermal growth factor (EGF). CMDB competed with the FGF-2 receptor for binding to FGF-2 but did not disturb the binding of IGF-1 and EGF to their receptors. Thus, our results highlight the selectivity of CMDB and their fragments towards FGF-2. Heparin, however, competes with CMDB and their fragments for binding to FGF-2. The carboxymethyl and benzylamide groups of these molecules likely interact directly with a heparin-binding region of FGF-2. The resulting change in conformation disturbs the binding of FGF-2 to its receptor and consecutively its mitogenic activity.

Sulphated polysaccharides derived from dextran: biomaterials for vascular therapy

CMDBS are synthetic dextran derivatives randomly substituted with carboxymethyl (CM), benzylamide (B), sulphonate and sulphate groups (S). Depending on their overall composition, these compounds are endowed with heparin-like properties such as anticoagulant activity. Indeed, some CMDBS with high CM and S contents delay blood coagulation, whilst some derivatized dextrans without significant anticoagulant capacity are potent antiproliferative agents for rat smooth muscle cells (SMCs) in vitro as well as heparin. The growth inhibition is dose dependent, reversible and nontoxic. This result is of prime interest for medical use because proliferation of vascular SMCs is postulated to be a key step in the pathogenesis of atherosclerosis or restenosis after vascular surgery such as angioplasty. By varying the overall composition in the different substituents, we have also prepared CMDBS exhibiting a stimulatory effect on the in vitro growth of human endothelial cells (EC). Heparin, under similar experimental conditions, slightly inhibited EC growth. The data indicate a synergistic role of all substituents grafted onto the dextran backbone without considering that any can be responsible alone for this effect. We conclude that a suitable distribution of CM, B and S groups on dextran can mimic heparin activity in terms of anticoagulant activity and antiproliferative capacity on SMC growth. Moreover, some CMDBS are also endowed with a stimulatory effect on EC growth. These properties confer great interest to these synthetic polysaccharides for vascular therapy.

Comparative anticoagulant activity and influence on thrombin generation of dextran derivatives and of a fucoidan fraction

CMDBS compounds are synthetic dextran derivatives with a random distribution of glucosylunits substituted with carboxymethyl, benzylamide, sulfonate, and sulfate groups. Fucoidans are sulfatedpolysaccharides extracted from brown seaweeds. CMDBS and fucoidans exhibit anticoagulant activitywhich depends on their chemical composition and molecular weight. Tested with purified proteins,these compounds catalyse thrombin (EC 3.4.21.5) inhibition mainly via heparin cofactor II (HCII). Weinvestigated the mechanism involved in the anticoagulant activity of these polysaccharides relative to thatof heparin. Three CMDBS with different chemical compositions were studied to evaluate the effect ofsulfate and sulfonate groups on the anticoagulant activity. The fucoidan fraction was extracted fromthe brown seaweed Ascophylum nodosum. The clotting assays (activated partial thromboplastin time,thrombin time, prothrombin time) were significantly prolonged in the presence of CMDBS and fucoidan,which were less active than heparin. To investigate the action mechanism of these polysaccharides,thrombin generation tests (TGT) were performed on human plasma in the presence of several CMDBSand a fucoidan fraction. The results showed an inhibition of thrombin generation in contact-activatedplasma in the presence of both polysaccharides, with a prolonged lag phase preceding the burst ofthrombin generation. In thromboplastin-activated plasma, thrombin generation was inhibited by CMDBSand fucoidan, with a prolonged lag phase only in the presence of CMDBS. The data obtained with eachpolysaccharide, compared to those obtained with heparin (our study) and hirudin (published data), ledto hypothesize that fucoidan could act, like heparin, by forming complexes with the inhibitor (althoughantithrombin (AT) in the case of heparin, and HCII for fucoidan), while CMDBS could act, like hirudin,by forming complexes with thrombin.

Heparin-like polymers derived from dextran enhance colonic anastomosis resistance to leakage.

A new tissue repair agent, RGTA11, is described for its ability to enhance colonic anastomosis repair and resistance to leakage. RGTA11 is a dextran derivative containing 110% carboxymethyl groups, 2.6% carboxymethyl benzylamide groups, and 36.6% carboxymethyl benzylamide sulfonate groups. RGTA11 was deemed efficient to protect the heparin-binding growth factors FGF2 against trypsin digestion. By this property RGTA11 mimicked heparin or heparan sulfate. We have also found that RGTA11 protected TGF beta 1 against trypsin digestion while heparin did not. RGTA11 was then tested in an in vivo wound-healing model of colonic anastomosis. Our results indicate that after 48 h, RGTA11- or RGTA11/FGF-2-treated animals presented a resistance of the anastomosis to leakage which was increased twofold (p < 0.05) over untreated controls. After 96 h and until day 7 there was no more difference with control animals. Our results suggest that RGTA11 presents potential clinical interest by preventing earlier leakage of colonic anastomosis.

Derivatized dextrans modulate collagen synthesis in aortic smooth muscle cells

The effect of specifically derivatized dextrans, with or without antiproliferative activity on smooth muscle cells (SMC), was investigated on type I and type III collagen biosyntheses and mRNA levels in post-confluent SMC cultures. Our results indicate that dextran derivatives decreased total protein and collagen syntheses independently of their antiproliferative activities. However, the most substituted dextran, the one exhibiting the strongest antiproliferative activity towards SMC, was the most active in modulating type III collagen expression. In addition, only the two dextran derivatives bearing benzylamide groups inhibited collagen excretion.

Functionalization of Poly(methyl acrylate). 1. Neighboring-Group Effects in the Reactions with Various Amines

An in situ carbon-13 NMR study of the kinetics and mechanism of the functionalization of poly(methyl acrylate) (PMA) by reactions with various amines is described. While the reaction with n-hexylamine (HAN) in a binary solvent of DMSO/o-dichlorobenzene (1:1, w/w) shows an autoacceleration effect, that with cyclohexanemethylamine (CN) exhibits autoretardation. The former is attributed to interactions that result in preferential solvation of nonpolar regions within the polymer domain, while the latter reflects the overwhelming effects of steric hindrance. Interestingly, while the presence of a single neighboring benzylamide group retards the reaction of benzylamine (BZA) with a PMA ester group, attack at an ester group flanked by benzylamide groups on either side is autoaccelerated

New Approaches to Tissue Regeneration and Repair

Several Heparin Binding Growth Factors (HBGFs) are thought to play a key role in the natural processes of tissue regeneration or repair after being released by neighbouring, inflammatory or circulating cells as well as from extracellular matrix associated heparan sulfate proteoglycosaminoglycans. In order to better understand how the bioavailability of these HBGFs can take part in the regulation of the wound healing processes, we have studied the healing effect of various chemically substituted dextrans (CMDBS) selected for their affinity for HBGFs, alone and in association with HBGFs. The CMDBS was obtained by substitution of methylcarboxylic (CM), benzylamide (B) and benzylamine sulfonate (S) groups in proportion of 83%, 23% and 13% respectively for CMDBS K that we have further used (Mauzac et al., 1985 Biomaterials. 6: 61-63). CMDBS K could 1: potentiate the biological activity of 1 or 2 FGFs, 2: protect 1 and 2 FGFs against thermal or pH inactivation, 3: protect a and b FGFs against proteolytic degradation (Tardieu et al., 1992 J. Cell. Physiol. 150: 194-203). CMDBS K was tested alone in cutaneous and bone wound healing models and for its ability to stabilize FGFs. Rats were punched and skin regeneration was studied by morphometric and histological analysis. The wounds (6 mm diameter) were filled with collagen plaster alone or soaked with CMDBS. CMDBS K in collagen plaster was able to induce a remarkable effect both on the kinetics and on the quality of the restored skin. These results suggest that endogenous growth factors naturally released during the regeneration process could be trapped, protected and released by CMDBS.(ABSTRACT TRUNCATED AT 250 WORDS)