Inhibitors of extracellular proteases

Abstract

Provided is a plant derived extract including inhibitory activity against one or more extracellular proteases which degrade human tissue matrix. Moreover, the amount of inhibitory activity in an extract can be increased by stressing the plant prior to forming an extract. These extracts are each prepared by a process and demonstrate the ability to inhibit one or more extracellular proteases which degrade human tissue matrix. Libraries of extracts can be prepared from stressed and non-stressed plants, where each of the extracts demonstrate inhibitory activity against on or more extracellular protease inhibitors. Alternatively, semi-purified and purified inhibitory compounds can be isolated from the extracts. In one aspect, these extracts with inhibitory activity can be used during protein purification to minimize degradation due to extracellular proteases

1 . A method of inhibiting an extracellular protease comprising contacting said extracellular protease with a plant extract, said plant extract capable of inhibiting the activity of one or more matrix metalloproteinase (MMP), one or more cathepsin, human leukocyte elastase (HLE), or a combination thereof, and said plant extract prepared by solvent extraction of plant material from  Aconitum napellus  L.;  Acorus calamus  L.;  Agrostis alba; Alchemilla mollis  L.;  Allium cepa  L.;  Allium porrum  L.;  Allium sativum  L.;  Allium schoenoporasum; Allium tuberosum; Aloe vera; Althaea officinalis  L.;  Amaranthus caudathus  L.;  Ambrosia artemisiifolia  L.;  Anethum graveolens  L.;  Angelica sinensis; Anthemis tinctoria  L.;  Arctostaphylos uva - ursi; Aronia melanocarpa  (Michx) Ell.;  Aronia×prunifolia; Artemisia dracunculus  L.;  Asarum europaeum; Avena sativa  L.;  Begonia mannii; Begonia polygonoides; Beta vulgaris  L.;  Beta vulgaris  L. subsp.  Maritima; Borago officinalis  L.;  Brassica napus  L.;  Brassica oleracea  L.;  Brassica rapa  L.;  Bromus inermis; Capsicum annuum  L. var.  annuum; Chaerophyllum bulbosum  L.;  Chenopodium quinoa  Willd.;  Cichorium endivia  L.;  Cichorium intybus; Circium arvense  L. Scop.;  Citrullus lanatus; Cornus canadensis  L.;  Daucus carota; Dioscorea batatas; Dirca palustris  L.;  Dolichos lablab  L.;  Fagopyrum esculentum  Moench;  Fagopyrum tartaricum; Foeniculum vulgare  Mill.;  Fragaria×ananassa; Frangula alnus  Mill.;  Gaultheria procumbens; Gentiana lutea  L.;  Geranium×cantabrigiense; Glycine max; Glycyrrhiza glabra  L.;  Hamamelis virginiana  L.;  Helianthus strumosus; Heliotropium arborescens  L.;  Hordeum vulgare  L. subsp.  vulgare; Humulus lupulus  L.;  Hypericum perforatum; Hypomyces lactiflorum; Iberis sempervirens; Juniperis communis  L.;  Kochia scoparia  L.;  Lactuca sativa  L.;  Lentinus edodes; Lotus corniculatus  L.;  Lunaria annua  L.;  Manihot esculenta; Matricaria recutita  L.;  Melilotus albus  Medik.;  Melilotis officinalis; Melissa officinalis; Mentha×piperita; Ocimum basilicum; Oenothera biennis  L.;  Origanum majorana  L.;  Origanum vulgare  L.;  Panax quinquefolius  L.;  Pastinaca sativa  L. subsp.  sativa; Petroselinum crispum; Phalaris canariensis  L.;  Phaseolus vulgaris  L. subsp.  vulgaris; Physostegia virginiana  L. Benth.;  Phytolacca decandra; Pimpinella anisum  L.;  Pisum sativum  L.;  Plantago major  L.;  Polygonum aviculare; Polygonum persicaria  L.;  Potentilla anserina  L.;  Poterium sanguisorba  L.;  Pyrus communis  L.;  Raphanus raphanistrum  L.;  Rheum rhabarbarum; Ribes nigrum  L.;  Ribes rubrum  L.;  Rodgersia  spp.;  Rosmarinus officinalis  L.;  Rubus allegheniensis; Rubus idaeus  L.;  Rubus occidentalis  L.;  Rubus thibetanus; Rumex crispus  L.;  Rumex scutatus  L.;  Ruta graveolens  L.;  Salvia officinalis  L.;  Sambucus canadensis  L.;  Sambucus ebulus  L.;  Saponaria officinalis  L.;  Satureja montana  L.;  Setaria italica; Silene vulgaris; Solanum melongena  L.;  Solidago canadensis; Solidago virgaureae; Solidago×hybrida; Sorghum caffrorum; Sorghum dochna; Stellaria media  (L.) Vill.;  Stephanandra incise; Tamarindus indica  L.;  Tanacetum cinerariifolium; Tanacetum vulgare  L.;  Taraxacum officinale; Teucrium chamaedrys  L.;  Thymus citriodorus; Thymus fragrantissimus; Thymus praecox  subsp  arcticus; Thymus pseudoanuginosus; Thymus vulgaris; Trifolium incarnatum  L.; X  Triticosecale  spp.;  Tropaeolum majus  L.;  Tsuga canadensis  L.;  Tsuga diversifolia; Vaccinium angustifolium  Aiton.;  Vigna unguiculata; Vinca minor; Vitis  sp.;  Zea mays  L. subsp.  mays  or  Zingiber officinale.    
     
     
         2 . The method according to  claim 1 , wherein said plant extract is an  Avena sativa  L.;  Capsicum annuum  L. var.  annuum; Chenopodium quinoa  Willd.;  Geranium .times. cantabrigiense; Glycine max; Helianthus strumosus; Iberis sempervirens; Juniperis communis  L.;  Lotus comiculatus  L.;  Melilotus albus  Medik.;  Pastinaca sativa  L. subsp.  sativa; Potentilla anserina  L.;  Rheum rhabarbarum; Rubus idaeus  L.;  Salvia officinalis  L.;  Sambucus canadensis  L.;  Saponaria officinalis  L.;  Solidago canadensis; Solidago virgaureae; Taraxacum officinale; Thymus citriodorus; Thymus vulgaris ; X  Triticosecale  spp.;  Tropaeolum majus  L.;  Tsuga canadensis  L.;  Vaccinium angustifolium  Aiton;  Zea mays  L. subsp.  mays  or  Zingiber officinale  extract. 
     
     
         3 . The method according to  claim 1 , wherein said extracellular protease is a MMP and said plant extract is an  Aconitum napellus  L.;  Acorns calamus  L.;  Agrostis alba; Alchemilla mollis  L.;  Allium cepa  L.;  Allium porrum  L.;  Allium sativum  L.;  Allium schoenoporasum; Allium tuberosum; Aloe vera; Althaea officinalis  L.;  Amaranthus caudathus  L.;  Ambrosia artemisiifolia  L.;  Anethum graveolens  L.;  Anthemis tinctoria  L.;  Arctostaphylos uva - ursi; Aronia melanocarpa  (Michx) Ell.;  Aronia .times. prunifolia; Artemisia dracunculus  L.;  Avena sativa  L.;  Beta vulgaris  L.;  Beta vulgaris  L. subsp.  Maritima; Borago officinalis  L.;  Brassica napus  L.;  Brassica oleracea  L.;  Brassica rapa  L.;  Bromus inermis; Capsicum annuum  L. var.  annuum; Chaerophyllum bulbosum  L.;  Chenopodium quinoa  Willd.;  Cichorium endivia  L.;  Circium arvense  L. Scop.;  Citrullus lanatus; Cornus canadensis  L.;  Daucus carota; Dioscorea batatas; Dirca palustris  L.;  Dolichos lablab  L.;  Fagopyrum esculentum  Moench;  Fagopyrum tartaricum; Foeniculum vulgare  Mill.;  Fragaria .times. ananassa; Gentiana lutea  L.;  Glycine max; Glycyrrhiza glabra  L.;  Hamamelis virginiana  L.;  Helianthus strumosus; Heliotropium arborescens  L.;  Hordeum vulgare  L. subsp.  vulgare; Humulus lupulus  L.;  Hypomyces lactiflorum; Juniperis communis  L.;  Kochia scoparia  L.;  Lactuca sativa  L.;  Lentinus edodes; Lotus comiculatus  L.;  Lunaria annua  L.;  Manihot esculenta; Matricaria recutita  L.;  Melilotus albus  Medik.;  Melilotis officinalis; Mentha×piperita; Oenothera biennis  L.;  Origanum majorana  L.;  Origanum vulgare  L.;  Panax quinquefolius  L.;  Pastinaca sativa  L. subsp.  sativa; Petroselinum crispum; Phalaris canariensis  L.;  Phaseolus vulgaris  L. subsp.  vulgaris; Phytolacca decandra; Pimpinella anisum  L.;  Pisum sativum  L.;  Plantago major  L.;  Polygonum persicaria  L.;  Potentilla anserina  L.;  Poterium sanguisorba  L.;  Pyrus communis  L.;  Raphanus raphanistrum  L.;  Rheum rhabarbarum; Ribes nigrum  L.;  Rosmarinus officinalis  L.;  Rubus allegheniensis; Rubus idaeus  L.;  Rubus occidentalis  L.;  Rubus thibetanus; Rumex crispus  L.;  Rumex scutatus  L.;  Ruta graveolens  L.;  Salvia officinalis  L.;  Sambucus canadensis  L.;  Sambucus ebulus  L.;  Saponaria officinalis  L.;  Satureja montana  L.;  Setaria italica; Silene vulgaris; Solanum melongena  L.;  Solidago canadensis; Solidago virgaureae; Sorghum caffrorum; Sorghum dochna; Stellaria media  (L.) Vill.;  Tanacetum cinerariifolium; Tanacetum vulgare  L.;  Taraxacum officinale; Teucrium chamaedrys  L.;  Thymus citriodorus; Thymus fragrantissimus; Thymus praecox  subsp  arcticus; Thymus pseudoanuginosus; Thymus vulgaris; Trifolium incarnatum  L.; X  Triticosecale  spp.;  Tropaeolum majus  L.;  Tsuga canadensis  L.;  Tsuga diversifolia; Vaccinium angustifolium  Aiton.;  Vigna unguiculata; Vinca minor; Vitis  sp.;  Zea mays  L. subsp.  mays  or  Zingiber officinale  extract. 
     
     
         4 . The method according to  claim 1 , wherein said extracellular protease is MMP-9 and said plant extract is an  Aconitum napellus  L.;  Acorns calamus  L.;  Agrostis alba; Alchemilla mollis  L.;  Allium cepa  L.;  Allium porrum  L.;  Allium sativum  L.;  Allium schoenoporasum; Allium tuberosum; Althaea officinalis  L.;  Amaranthus caudathus  L.;  Ambrosia artemisiifolia  L.;  Anthemis tinctoria  L.;  Arctostaphylos uva - ursi; Aronia melanocarpa  (Michx) Ell.;  Aronia .times. prunifola; Artemisia dracunculus  L.;  Avena sativa  L.;  Beta vulgaris  L.;  Beta vulgaris  L. subsp.  Maritima; Borago officinalis  L.;  Brassica napus  L.;  Brassica oleracea  L.;  Brassica rapa  L.;  Bromus inermis; Capsicum annuum  L. var.  annuum; Chaerophyllum bulbosum  L.;  Chenopodium quinoa  Willd.;  Cichorium endivia  L.;  Citrullus lanatus; Cornus canadensis  L.;  Daucus carota; Dioscorea batatas; Dirca palustris  L.;  Dolichos lablab  L.;  Fagopyrum esculentum  Moench;  Fagopyrum tartaricum; Foeniculum vulgare  Mill.;  Fragaria .times. ananassa; Gentiana lutea  L.;  Glycine max; Glycyrrhiza glabra  L.;  Hamamelis virginiana  L.;  Helianthus strumosus; Heliotropium arborescens  L.;  Hordeum vulgare  L.;  Humulus lupulus  L.;  Hypomyces lactiflorum; Juniperis communis  L.;  Lactuca sativa  L.;  Lentinus edodes; Lotus corniculatus  L.;  Lunaria annua  L.;  Manihot esculenta; Matricaria recutita  L.;  Melilotus albus  Medik.;  Meliotis officinalis; Mentha×piperita; Oenothera biennis  L.;  Origanum majorana  L.;  Origanum vulgare  L.;  Panax quinquefolius  L.;  Pastinaca sativa  L. subsp.  sativa; Phalaris canariensis  L.;  Phaseolus vulgaris  L.;  Phytolacca decandra; Pimpinella anisum  L.;  Pisum sativum  L.;  Plantago major  L.;  Polygonum persicaria  L.;  Potentilla anserina  L.;  Poterium sanguisorba  L.;  Pyrus communis  L.;  Raphanus raphanistrum  L.;  Rheum rhabarbarum; Ribes nigrum  L.;  Rosmarinus officinalis  L.;  Rubus idaeus  L.;  Rubus allegheniensis; Rubus occidentalis  L.;  Rumex crispus  L.;  Rumex scutatus  L.;  Ruta graveolens  L.;  Salvia officinalis  L.;  Sambucus canadensis  L.;  Sambucus ebulus  L.;  Satureja montana  L.;  Setaria italica; Solanum melongena  L.;  Solidago canadensis; Solidago virgaureae; Sorghum caffrorum; Sorghum dochna; Tanacetum cinerariifolium; Tanacetum vulgare  L.;  Taraxacum officinale; Teucrium chamaedrys  L.;  Thymus citriodorus; Thymus fragrantissimus; Thymus pseudoanuginosus; Thymus vulgaris; Trifolium incarnatum  L.; X  Triticosecale  spp.;  Tropaeolum majus  L.;  Tsuga canadensis  L.;  Tsuga diversifolia; Vaccinium angustifolium  Aiton.;  Vigna unguiculata; Vinca minor; Vitis  sp.;  Zea mays  L. subsp.  mays  or  Zingiber officinale  extract. 
     
     
         5 . The method according to  claim 1 , wherein said extracellular protease is MMP-9 and said plant extract is an  Avena sativa  L.;  Capsicum annuum  L. var.  annuum; Chenopodium quinoa  Willd.;  Glycine max; Helianthus strumosus; Juniperis communis  L.;  Lotus comiculatus  L.;  Melilotus albus  Medik.;  Pastinaca sativa  L. subsp.  sativa; Potentilla anserina  L.;  Rheum rhabarbarum; Rubus idaeus  L.;  Salvia officinalis  L.;  Sambucus canadensis  L.;  Solidago canadensis; Solidago virgaureae; Taraxacum officinale; Thymus citriodorus; Thymus vulgaris ; X  Triticosecale  spp.;  Tropaeolum majus  L.;  Tsuga canadensis  L.;  Vaccinium angustifolium  Aiton;  Zea mays  L. subsp.  mays  or  Zingiber officinale  extract. 
     
     
         6 . The method according to  claim 1 , wherein said extracellular protease is cathepsin B and said plant extract is an  Aconitum napellus  L.;  Alchemilla mollis  L.;  Allium cepa  L.;  Allium sativum  L.;  Allium schoenoporasum; Allium tuberosum; Althaea officinalis  L.;  Angelica sinensis; Anthemis tinctoria  L.;  Arctostaphylos uva - ursi; Aronia melanocarpa  (Michx) Ell.;  Aronia .times. prunifolia; Artemisia dracunculus  L.;  Asarum europaeum; Begonia mannii; Begonia polygonoides; Beta vulgaris  L.;  Beta vulgaris  L. subsp.  Maritima; Borago officinalis  L.;  Brassica napus  L.;  Brassica oleracea  L.;  Brassica rapa  L.;  Capsicum annuum  L. var.  annuum; Citrullus lanatus; Daucus carota; Dirca palustris  L.;  Fragaria .times. ananassa; Glycine max; Glycyrrhiza glabra  L.;  Hamamelis virginiana  L.;  Heliotropium arborescens  L.;  Humulus lupulus  L.;  Hypomyces lactiflorum; Iberis sempervirens; Kochia scoparia  L.;  Lentinus edodes; Lunaria annua  L.;  Origanum majorana  L.;  Origanum vulgare  L.;  Panax quinquefolius  L.;  Pastinaca sativa  L. subsp.  sativa; Petroselinum crispum; Phaseolus vulgaris  L.;  Physostegia virginiana  L. Benth.;  Phytolacca decandra; Plantago major  L.;  Ribes nigrum  L.;  Rubus allegheniensis; Rubus idaeus  L.;  Rubus occidentalis  L.;  Rumex crispus  L.;  Salvia officinalis  L.;  Sambucus canadensis  L.;  Saponaria officinalis  L.;  Satureja montana  L.;  Silene vulgaris; Solidago canadensis; Solidago .times. hybrida; Sorghum caffrorum; Tamarindus indica  L.;  Tanacetum vulgare  L.;  Taraxacum officinale; Thymus praecox  subsp  arcticus; Thymus vulgaris; Tropaeolum majus  L.;  Tsuga diversifolia; Vaccinium angustifolium  Aiton.;  Vitis  sp. or  Zea mays  L. subsp.  mays  extract. 
     
     
         7 . The method according to  claim 1 , wherein said extracellular protease is cathepsin B and said plant extract is a  Capsicum annuum  L. var.  annuum; Glycine max; Iberis sempervirens; Pastinaca sativa  L. subsp.  sativa; Rubus idaeus  L.;  Salvia officinalis  L.;  Sambucus canadensis  L.;  Saponaria officinalis  L.;  Solidago canadensis; Taraxacum officinale; Thymus vulgaris; Tropaeolum majus  L.;  Vaccinium angustifolium  Aiton. or  Zea mays  L. subsp.  mays  extract. 
     
     
         8 . The method according to  claim 1 , wherein said extracellular protease is HLE and said plant extract is an  Aconitum napellus  L.;  Acorns calamus  L.;  Alchemilla mollis  L.;  Allium sativum  L.;  Allium tuberosum; Aloe vera; Althaea officinalis  L.;  Amaranthus caudathus  L.;  Anethum graveolens  L.;  Anthemis tinctoria  L.;  Arctostaphylos uva - ursi; Aronia melanocarpa  (Michx) Ell.;  Aronia .times. prunifolia; Artemisia dracunculus  L.;  Avena sativa  L.;  Begonia mannii; Begonia polygonoides; Beta vulgaris  L.;  Beta vulgaris  L. subsp.  Maritima; Borago officinalis  L.;  Brassica napus  L.;  Brassica oleracea  L.;  Brassica rapa  L.;  Bromus inermis; Capsicum annuum  L. var.  annuum; Chaerophyllum bulbosum  L.;  Chenopodium quinoa  Willd.;  Cichorium endivia  L.;  Circium arvense  L. Scop.;  Citrullus lanatus; Cornus canadensis  L.;  Daucus carota; Dirca palustris  L.;  Dolichos lablab  L.;  Fagopyrum esculentum  Moench;  Fagopyrum tartaricum; Foeniculum vulgare  Mill.;  Fragaria .times. ananassa; Frangula alnus  Mill.;  Gentiana lutea  L.;  Geranium .times. cantabrigiense; Glycine max; Glycyrrhiza glabra  L.;  Hamamelis virginiana  L.;  Heliotropium arborescens  L.;  Hordeum vulgare  L.;  Humulus lupulus  L.;  Hypomyces lactiflorum; Iberis sempervirens; Juniperis communis  L.;  Lactuca sativa  L.;  Lentinus edodes; Matricaria recutita  L.;  Mentha×piperita; Oenothera biennis  L.;  Origanum majorana  L.;  Origanum vulgare  L.;  Pastinaca sativa  L. subsp.  sativa; Petroselinum crispum; Phalaris canariensis  L.;  Phaseolus vulgaris  L.;  Phytolacca decandra; Pimpinella anisum  L.;  Plantago major  L.;  Polygonum persicaria  L.;  Poterium sanguisorba  L.;  Ribes nigrum  L.;  Ribes rubrum  L.;  Rodgersia  spp.;  Rosmarinus officinalis  L.;  Rubus idaeus  L.;  Rubus occidentalis  L.;  Rubus thibetanus; Rumex crispus  L.;  Rumex scutatus  L.;  Ruta graveolens  L.;  Salvia officinalis  L.;  Sambucus canadensis  L.;  Sambucus ebulus  L.;  Satureja montana  L.;  Solanum melongena  L.;  Sorghum caffrorum; Sorghum dochna; Stellaria media  (L.) Vill.;  Stephanandra incise; Tanacetum cinerariifolium; Tanacetum vulgare  L.;  Taraxacum officinale; Teucrium chamaedrys  L.;  Thymus citriodorus; Thymus fragrantissimus; Thymus praecox  subsp  arcticus; Thymus pseudoanuginosus; Thymus vulgaris ; X  Triticosecale  spp.;  Tropaeolum majus  L.;  Tsuga diversifolia; Vaccinium angustifolium  Aiton.;  Vitis  sp. or  Zea mays  L. subsp.  mays  extract. 
     
     
         9 . The method according to  claim 1 , wherein said extracellular protease is HLE and said plant extract is an  Avena sativa  L.;  Capsicum annuum  L. var.  annuum; Chenopodium quinoa  Willd.;  Geranium .times. cantabrigiense; Glycine max; Iberis sempervirens; Juniperis communis  L.;  Pastinaca sativa  L. subsp.  sativa; Rubus idaeus  L.;  Salvia officinalis  L.;  Sambucus canadensis  L.;  Taraxacum officinale; Thymus citriodorus; Thymus vulgaris ; X  Triticosecale  spp.;  Tropaeolum majus  L.;  Vaccinium angustifolium  Aiton. or  Zea mays  L. subsp.  mays  extract.