DE69717133T2 - LIQUID DETERGENTS CONTAINING PROTEOLYTIC ENZYME, PEPTIDALDEHYDE AND CALCIUM IONS - Google Patents

Description

Technical area

This invention relates to enzyme-containing liquid detergent compositions. More particularly, this invention relates to liquid detergent compositions containing a detersive surfactant, a proteolytic enzyme, a peptide aldehyde, and calcium ions. The combination of peptide aldehyde and calcium ions serves to provide synergistic protease inhibitor benefits.

Background of the invention

Protease-containing liquid aqueous detergents are well known, particularly in the context of laundry washing. A common problem with such protease-containing liquid aqueous detergents is the degradation phenomenon of second enzymes in the composition, such as amylase, lipase and cellulase, by the proteolytic enzyme or the protease itself. As a result, the stability of the second enzyme or the protease itself in the detergent composition is impaired and the detergent composition is consequently less effective.

In response to this problem, the use of various protease inhibitors or stabilizers has been proposed. For example, several references have suggested the use of the following compounds to assist in the stabilization of enzymes: benzamidine hydrochloride, lower aliphatic alcohols or carboxylic acids, mixtures of a polyol and a boron compound, aromatic borate esters, and calcium, particularly calcium formate. Recently, it has been discovered that certain peptide aldehydes are effective in stabilizing the protease enzyme.

Although these compounds have been used with varying degrees of success in liquid detergents, they are not without problems. For example, peptide aldehydes are quite expensive and create complexities for formulators, especially for liquid detergents. Other inhibitors such as calcium and boric acids are cheaper but do not stabilize enzymes as well as peptide aldehydes. It is therefore an object of the present invention to provide a protease inhibitor system which is economical, effective and suitable for use in a liquid detergent composition.

In response to this aim, the present invention proposes the use of a combination of calcium ions and peptide aldehydes as reversible protease inhibitors in aqueous liquid detergent compositions. The presence of both calcium and a peptide aldehyde provides synergistic stabilization of the protease. This new combination provides the formulator with more flexibility in developing a stabilization system. The proportions of peptide aldehyde and calcium can be adjusted to provide the most cost-effective formula and to minimize product stability problems that often arise from the presence of divalent ions in a liquid detergent matrix.

In particular, the present invention enables the use of very low levels of peptide aldehydes in the liquid detergent compositions herein. This is particularly important in the formulation of relatively inexpensive concentrated liquid detergent compositions, which are included in the present invention.

Because the combination of calcium and peptide aldehydes is so effective in inhibiting proteases, another advantage of the present invention is that even enzymes that are particularly sensitive to proteolytic degradation can now be incorporated into liquid detergent compositions comprising a protease. In addition, it has also been recognized that the increased stability of the protease enzyme allows for improved skin care benefits. These benefits include softening of the skin and hands, as well as reduced drying from exposure of the hands to dishwashing detergent solution.

Background of the field

The use of various protease inhibitors or stabilizers has been proposed. For example, US 4,566,985 suggests using benzamidine hydrochloride; EP 376 705 suggests using lower aliphatic alcohols or carboxylic acids; EP 381 262 suggests using a mixture of a polyol and a boron compound; and EP 91870072.5 suggests using aromatic borate esters. See also US Patent No. 5,030,378, issued July 9, 1991. See also US 4,261,868; US 4,404,115; US 4,318,818; and EP 130,756.

The use of peptide derivatives to inhibit proteins appears to have been disclosed in therapeutic applications. EP 293 881 discloses the use of peptide boronic acids as inhibitors of trypsin-like serine proteases. EP 185 390 and US 4,399,065 disclose the use of certain peptide aldehyde derivatives to inhibit blood coagulation. J-90029670 generally discloses the use of optically active alpha-amino aldehydes to inhibit enzymes. See also "Inhibition of Thrombin and Trypsin by Tripeptide Aldehydes", Int. J. Peptide Protein Res., vol. 12 (1978), pp. 217-221, Gaal, Bacsy &Rappay; and "Tripeptide Aldehyde Protease Inhibitors May Depress in Vitro Prolactin and Growth Hormone Release", Endocrinology, Vol. 116, No. 4 (1985), pp. 1426-1432, Rappay, Makara, Bajusz & Nagy. Certain peptide aldehydes have also been disclosed in EP-A-473 502 for inhibiting protease-mediated skin irritation.

See in particular EP 185,390; WO 94/04651, published March 3, 1994; WO 94/04652, published March 3, 1994; EP 583,536, published February 23, 1994; EP 583,535, published February 3, 1994; EP 583,534, published February 23, 1994; WO 93/13125, published July 8, 1993; US 4,529,525; US 4,537,706; US 4,537,707; and US 5,527,487.

JP 62269689 describes a method for stabilizing enzymes in detergent compositions by adding a reversible inhibitor for the enzyme and optionally adding a calcium salt. WO 92/03529 discloses a detergent composition comprising a protease and one or more enzymes and a reversible protease inhibitor of the peptide or protein type.

Summary of the invention

The invention herein is a liquid detergent composition comprising:

(a) an effective amount of a detersive surfactant;

b) an active proteolytic enzyme;

c) a source of calcium ions other than a compound of the formula RO(A)mSO₃M, wherein R, A and M are as described in claim 1; and

d) a peptide aldehyde of the formula:

Z-B-NH-CH(R)-C(O)H

wherein B is a peptide chain comprising 1 to 5 amino acid groups; Z is an N-capping group selected from the group consisting of phosphoramidate [(R"O)₂(O)P-], sulfenamide [(SR")₂-], sulfonamide [(R"(O)₂S-], sulfonic acid [SO₃H], phosphinamide [(R")₂(O)P-], sulfamoyl derivative [R"O(O)₂S-], thiourea [(R")₂N(O)C-], thiocarbamate [R"O(S)C-], phosphonate [R"-P(O)OH], amidophosphate [R"O(OH)(O)P-], carbamate (R"O(O)C-) and urea (R"NH(O)C-), wherein each R" is independently selected from the group consisting of straight or branched, unsubstituted C₁-C₆ alkyl, phenyl, C₇-C₆ alkylaryl and cycloalkyl groups, wherein the cycloalkyl ring may span C₄-C₈ and may contain one or more heteroatoms, selected from the group consisting of O, N and S (preferably R" is selected from the group consisting of methyl, ethyl and benzyl); and R is selected from the group consisting of straight or branched, unsubstituted C₁-C₆ alkyl, phenyl and C₇-C₆ alkylaryl groups.

Without being bound to any theory, it is believed that the combination of calcium ion source and peptide aldehyde provides more than additive stability for the proteolytic enzyme.

Preferably, the liquid detergent compositions herein comprise by weight of the composition:

a) 1 to 95%, preferably 8 to 70%, of the detergent surfactant;

b) 0.0001 to 5%, preferably 0.0003 to 0.1%, of an active proteolytic enzyme;

c) 0.00001 to 5%, preferably 0.0001 to 1%, more preferably 0.0006 to 0.5%, of a peptide aldehyde as described above; and

d) 0.01 to 1%, preferably 0.05 to 0.5%, of calcium ions.

The proteolytic enzyme useful herein is preferably a subtilisin-type protease and may be selected from the group consisting of Alcalase®, Subtilisin BPN', Protease A, Protease B and mixtures thereof.

The calcium ion source for use herein is preferably selected from calcium formate, calcium xylene sulfonate, calcium chloride, calcium acetate, calcium sulfate, and mixtures thereof.

The dish care compositions herein may contain additional washing additives, including, but not limited to, one or more of the following: suds boosters, chelating agents, polyacrylate polymers, dispersants, dyes, perfumes, processing aids, and mixtures thereof. In addition, the liquid detergent compositions for dish care compositions may further comprise an effective amount of amylase enzyme. In addition, the dish care compositions may optionally comprise an effective amount of a source of boric acid and a diol. Typical dish care compositions optionally, but preferably, comprise from about 0.25 to about 10%, preferably from about 0.5 to about 5%, more preferably from about 0.75 to about 3%, by weight of boric acid or a compound capable of forming boric acid and a diol, e.g., 1,2-propanediol.

In a preferred embodiment, the liquid detergent composition for general purpose detergent compositions further comprises an effective amount of one or more of the following enzymes: lipase, amylase, cellulase, and mixtures thereof. For detergent compositions, the second enzyme is preferably a lipase and is obtained by cloning the gene from Humicola lanuginosa and expressing the gene in Aspergillus oryzae. Lipase is used in an amount of from about 10 to about 18,000 lipase units per gram, preferably from about 60 to about 6,000 units per gram.

In another preferred composition useful for laundry care, the second enzyme is a cellulase derived from Humicola insolens and is used in an amount of about 0.0001 to about 0.1% by weight of the total cellulase composition.

The compositions herein may further contain laundry additives including, but not limited to, one or more of the following: foam boosters, builders, soil release polymers, polyacrylate polymers, dispersants, dye transfer inhibitors, dyes, perfumes, processing aids, brighteners, and mixtures thereof. In addition, the detersive surfactant for laundry care compositions is typically present in an amount of 10 to 70% by weight of the total composition. Additionally, the detergent compositions may optionally comprise an effective amount of a source of boric acid and a diol. Typical detergent compositions optionally, but preferably, comprise from about 0.25 to 10% by weight, preferably from about 0.5 to about 5% by weight, more preferably from about 0.75 to about 3% by weight, of boric acid or a compound capable of forming boric acid and a diol, e.g., 1,2-propanediol.

All percentages and ratios herein are by weight, and all references cited are hereby incorporated by reference unless otherwise specifically indicated.

Detailed description of the invention

Definitions - The present detergent compositions comprise an "effective amount" or a "stain removal enhancing amount" of individual components as defined herein. An "effective amount" or a "stain removal enhancing amount" is any amount capable of measurably improving soil cleaning or stain removal from a substrate, i.e., a soiled fabric or soiled dish, when washed or rinsed by the consumer. In general, this amount can vary widely.

By "synergy" or "more than additive" as used herein, it is meant that the enzyme stability benefit of combining calcium and peptide aldehydes is greater than the sum of the individual benefits obtained when only one of the components is present in a detergent composition.

The liquid aqueous detergent compositions of the present invention comprising four essential ingredients: (A) a peptide aldehyde or mixture thereof, (B) a proteolytic enzyme or mixture thereof, (C) a detersive surfactant, and (D) calcium ions. The compositions of the present invention preferably further comprise (E) a detergent-compatible second enzyme or mixture thereof, and may further comprise (F) other optional ingredients.

Peptide aldehydes

The detergent compositions according to the invention comprise as a first essential ingredient a peptide aldehyde having the formula:

Z-B-NH-CH(R)-C(O)H

wherein B is a peptide chain comprising 1 to 5 amino acid groups; Z is an N-capping group selected from the group consisting of phosphoramidate [(R"O)₂(O)P-], sulfenamide [(SR")₂-], sulfonamide [(R"(O)₂S-], sulfonic acid [SO₃H], phosphinamide [(R")₂(O)P-], sulfamoyl derivative [R"O(O)₂S-], thiourea [(R")₂N(O)C-], thiocarbamate [R"O(S)C-], phosphonate [R"-P(O)OH], amidophosphate [R"O(OH)(O)P-], carbamate (R"O(O)C-) and urea (R"NH(O)C-), wherein each R" is independently selected from the group consisting of straight or branched, unsubstituted C₁-C₆- Alkyl, phenyl, C₇-C₉ alkylaryl and cycloalkyl groups, wherein the cycloalkyl ring may span C₄-C₈ and may contain one or more heteroatoms, is selected from the group consisting of O, N and S (preferably R" is selected from the group consisting of methyl, ethyl and benzyl); and R is selected from the group consisting of straight or branched, unsubstituted C₁-C₆ alkyl, phenyl and C₇-C₉ alkylaryl groups.

Preferred R groups are selected from the group consisting of methyl, iso-propyl, sec-butyl, iso-butyl, -C6H5, -CH2-C6H5 and -CH2CH2-C6H5, each of which may be derived from the amino acids Ala, Val, Ile, Leu, PGly (phenylglycine), Phe and HPhe (homophenylalanine) by converting the carboxylic acid group to an aldehyde group. Although such groups are therefore not amino acids (and they may or may not have been synthesized from an amino acid precursor), for the purpose of simplifying the illustration of inhibitors useful herein, the aldehyde moiety of the inhibitors is indicated as being derived from amino acids by appending an "H" to the analogous amino acid [e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 49, 53, 54, 55, 56, 57, 58, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 80, 81, 82, 83, 84 For example, "-AlaH" means the chemical group "-NHCH(CH3)C(O)H".

Preferred B-peptide chains are selected from the group consisting of peptide chains having the amino acid sequences according to the general formula:

Z-A⁵-A⁴-A³-A²-A¹-NH-CH(R)-C(O)H

so that the following amino acids, if present, are:

A¹ selected from Ala, Gly;

A², if available, chosen from Val, Ala, Gly, Ile;

A³, if present, selected from Phe, Leu, Val, Ile;

A⁴, if present, any amino acid, but is preferably selected from Gly, Ala;

A⁵, if present, any amino acid, but is preferably Gly, Ala, Lys.

The aldehydes of the invention can be prepared from the corresponding amino acid, whereby the C-terminal end of the amino acid is converted from a carboxylic acid group to an aldehyde group. Such aldehydes can be prepared by known methods, for example as described in US 5015627, EP 185 390, EP 583,534 and DE 32 00 812.

Without being bound by theory, it is believed that the peptide aldehydes of the invention bind to the proteolytic enzyme in the liquid detergent composition, thereby inhibiting the proteolytic enzyme. After dilution with water, the proteolytic enzyme is returned to its original state by dissociation of the proteolytic enzyme/peptide aldehyde complex.

The N-terminal end of the protease inhibitors according to the invention is protected by one of the N-capping group protecting groups selected from the group consisting of carbamates, ureas, sulfonamides, phosphonamides, thioureas, sulfenamides, sulfonic acids, phosphinamides, thiocarbamates, amidophosphates and phosphonamides. However, in a particularly preferred embodiment of the present invention, the N-terminal end of the protease inhibitor is protected by a methyl, ethyl or benzyl carbamate [CH₃O�min;(O)C-; CH₃CH₂O-(O)C-; or C₆H₅CH₂O-(O)C-], methyl, ethyl or benzyl urea [CH₃NH-(O)C-; CH₃CH₂NH-(O)C-; or C₆H₅CH₂NH-(O)C-], methyl, ethyl or benzyl sulfonamide [CH₃SO₂-; CH₃CH₂SO₂-; or C₆H₅CH₂SO₂-] and methyl, ethyl or benzyl amidophosphate [CH₃O(OH)(O)P-; CH₃CH₂O(OH)O(P)-; or C₆H₅CH₂O(OH)(O)P-] group.

The synthesis of N-capping groups can be found in the following references: Protective Groups in Organic Chemistry, Greene T., Wuts P., John Wiley & Sons, New York, 1991, pp. 309-405; March J., Advanced Organic Chemistry, Wiley Interscience, 1985, pp. 445, 469; Carey F., Sundberg R., Advanced Organic Chemistry, Part B, Plenum Press, New York, 1990, pp. 686-689; Atherton E., Sheppard R., Solid Phase Peptide Synthesis, Pierce Chemical, 1989, pp. 3-4; Grant G., Synthetic Peptides, W. H. Freeman & Co., 1992, pp. 77-103; Stewart J., Young J., Solid Phase Peptide Synthesis, 2nd edition, IRL Press, 1984, pp. 3, 5, 11, 14-18, 28-29; Bodansky M., Principles of Peptide Synthesis, Springer-Verlag, 1988, pp. 62, 203, 59-69; Bodansky M., Peptide Chemistry, Springer-Verlag, 1988, pp. 74-81; Bodansky M., Bodansky A., The Practice of Peptide Synthesis, Springer-Verlag, 1984, pp. 9-32.

Examples of peptide aldehydes for use herein are:

CH₃SO₂Phe-Gly-Ala-Leu-H, CH₃SO₂Val-Ala-Leu-H, C₆H₅CH₂O(OH)(O)P-Val-Ala-Leu-H, CH₃CH₂SO₂- Phe-Gly-Ala-Leu-H, C₆H₅CH₂SO₂-Val-Ala-Leu-H, C₆H₅CH₂O(OH)(O)P-Leu- Ala-Leu-H, C₆H₅CH₂O(OH)(O)P-Phe-Ala-Leu-H and CH3 O(OH)(O)P-Leu-Gly-Ala-Leu-H.

In the following synthesis examples, methods for synthesizing certain of these peptide aldehydes are disclosed.

Synthesis example 1 Synthesis of the tetrapeptide aldehyde Moc-Ala-Phe-Gly-Ala-LeuH

(a) Ala-Leu-OMe·HCl: To a solution of 3.0 g (14.83 mmol) of Ala-Leu-OH dissolved in 50 mL of MeOH and cooled to 0ºC is added dropwise 2.43 mL (33.36 mmol) of thionyl chloride. This solution is stirred overnight at room temperature and evaporated to dryness to provide quantitative recovery of the desired product.

(b) Cbz-Gly-Ala-leucine methyl ester: To a solution of 0.414 g (1.98 mmol) of Cbz-Gly-OH and 0.500 g (1.98 mmol) of Ala-Leu-OMe·HCl in CH2Cl2 is added 0.607 mL of TEA, immediately followed by 0.355 mL of DEPC. The solution is stirred overnight, evaporated, and the residue is partitioned between EtOAc and 1 N HCl. The organic layer is washed sequentially with saturated NaHCO3. and saturated NaCl, dried (MgSO4) and evaporated to give 0.650 g of the crude product.

(c) Moc-Ala-Phe-OH; To a solution of 1.0 g (4.23 mmol) of Ala-Phe dissolved in 4.23 mL of 1 N NaOH and cooled to 0ºC is added dropwise 0.419 g (4.44 mmol) of methyl chloroformate. Simultaneously, in a separate addition funnel, 4.23 mL of 1 N NaOH is added so that the pH is maintained between 9.0-9.5. After the addition is complete, the reaction is stirred for 30 min at 0ºC and 2 h at room temperature. At this time, the solution is cooled to 0ºC and the pH is adjusted to 9.5. This basic solution is washed with EtOAc (1x, 100 mL). The aqueous phase (0°C) is then adjusted to pH = 2.5 (2 N HCl) and extracted with EtOAc (3x, 50 mL), dried (MgSO4) and evaporated to provide 1.07 g of the crude product.

(d) Moc-Ala-Phe-Gly-Ala-Leu-OMe: To a solution of 0.500 g (1.22 mmol) of Cbz- Gly-Ala-leucine methyl ester in 10 mL of MeOH is added 0.100 g of 10% Pd/C. This solution is hydrogenated in the presence of 0.600 mL of 4.0 M HCl/dioxane (under balloon pressure) for 1 h, filtered through celite and evaporated. This residue is suspended in CH2Cl2, 0.342 mL (2.45 mmol) of TEA is added, followed by 0.359 g (1.22 mmol) of Moc-Ala-Phe-OH and 0.219 mL (1.34 mmol) of DEPC. After stirring overnight, the solvent is evaporated, the residue is partitioned between EtOAc and 1 N HCl and washed sequentially with saturated NaHCO3 and NaCl. Drying, evaporation and column chromatography give 0.450 g of the crude product.

(e) Moc-Ala-Phe-Gly-Ala-Leucinol: A solution is prepared by dissolving 0.182 g (1.64 mmol) of CaCl2 in a mixture of 4 mL of ethanol and 2 mL of THF. This mixture is cooled to -15 °C and 0.450 g (0.820 mmol) of Moc-Ala-Phe- Gly-Ala-Leu-OMe is added followed by 0.124 g (3.28 mmol) of NaBH4. The reaction is stirred for 2 h and quenched with 10 mL of 1 N HCl. The solvents are evaporated and the remaining aqueous layer is partitioned with EtOAc. The organic phase is then washed with saturated NaHCO3 and saturated NaCl. Drying (MgSO4), evaporation and chromatography give 0.256 g of the crude product.

(1) Moc-Ala-Phe-Gly-Ala-LeuH: A solution is prepared by adding 0.623 g (1.47 mmol) of Dess-Martin periodinane to 1.8 L of CH2Cl2, followed by stirring for 10 min. This solution is then cooled to 0 °C and 0.256 g (0.490 mmol) of Etoc-Phe-Gly-Ala-leucinol is added in one portion. The reaction is continued for 2 h and poured into a solution consisting of 2.55 g (10.47 mmol) of Na2S2O3 in 30 mL of saturated NaHCO3. After stirring for 10 min, the mixture is extracted with EtOAc (2x, 50 mL). The combined extracts are dried (MgSO4), evaporated and chromatographed on silica to provide 0.125 g of the crude product.

Synthesis example 2 Synthesis of the tripeptide aldehyde Etoc-Phe-Gly-Ala-LeuH

(a) Ala-Leu-OMe·HCl: To a solution of 450 g (2.20 mol) of Ala-Leu-OH dissolved in 4.5 L of MeOH and cooled to 0ºC is added dropwise 178.6 mL (4.95 mol) of thionyl chloride. The solution is stirred overnight at room temperature and evaporated to dryness to provide 543 g (97.1% yield) of the desired product, which is to be used as such.

(b) Etoc-Phe-Gly-OH: To a solution of 450 g (2.03 mol) of Phe-Gly dissolved in 2026 mL of 1 N NaOH and cooled to 0ºC is added dropwise methyl chloroformate (3.1 mL, 40.0 mmol). Simultaneously, in a separate addition funnel, an additional 2026 mL of 1 N NaOH is added so that the pH is maintained between 9.0-9.5. After the addition is complete, the reaction is stirred for 30 min at 0ºC and 2 h at room temperature. At this time, the solution is cooled to 0ºC and the pH is adjusted to 9.5. This basic solution is washed with EtOAc (1x, 4 L). The aqueous phase (0 °C) is then adjusted to pH = 2.5 (2 N HCl) and extracted with EtOAc (3x, 8 L), dried (MgSO4), filtered and the solvent removed to give 546 g (91.3% yield) of the crude product.

(c) Etoc-Phe-Gly-Ala-Leu-OMe: To a solution of 470 g (1.86 mol) of Etoc-Phe-Gly-OH and 546 g (1.86 mol) of Ala-Leu-OMe·HCl in 8 L of CH2Cl2 is added 570 mL (4.09 mol) of TEA followed by 310.4 mL (2.046 mol) of DEPC. After stirring overnight, the solvent is evaporated and replaced with EtOAc (4 L). This solution is washed sequentially with 2 L each of 2 N HCl, saturated NaHCO3 and saturated NaCl. The organic layer is then dried (MgSO4), filtered and evaporated to give 916 g (93% yield) of the desired material.

(d) Etoc-Phe-Gly-Ala-Leucinol: To a solution of 45.10 g (0.406 mol) of CaCl2 in 1 L ethanol and 1 L THF is added 100 g (0.203 mol) of Etoc-Phe-Gly-Ala-Leu-OMe and the mixture is cooled to -15 °C. To this solution is carefully added 30.7 g (0.812 mmol) of NaBH4 followed by stirring for 2 h. Then the reaction is quenched with 100 mL of 0.1 N HCl. This solution is transferred to 4 L of 1 N HCl and extracted with EtOAc (3x, 2.75 L). The combined EtOAc layers are washed with 4 L of saturated NaHCO3. washed, dried (MgSO4) and evaporated. Trituration (twice) with ether (4 L) afforded 69.2 g (73.4% yield) of the product.

(e) Etoc-Phe-Gly-Ala-LeuH: A solution is prepared by adding 165.4 g (0.39 mol) of Dess-Martin periodinane to 1.8 L of CH2Cl2, followed by stirring for 10 minutes. This solution is then cooled to 0°C, and 60 g (0.13 mol) of Etoc-Phe-Gly-Ala-leucinol is added in one portion. The reaction is continued for 105 minutes and poured into a solution consisting of 6 L of H2O, 393 g of NaHCO3, and 431.7 g (1.74 mol) of Na2S2O3. After stirring for 10 min, the phases are separated and two more extractions (1.5 L each) with CH2Cl2 are carried out. The combined extracts are dried (MgSO4), filtered and triturated with ether (2x, 1 L) to provide 51.7 g (86.2% yield) of the product.

Synthesis example 3 Synthesis of the dipeptide aldehyde Moc-Gly-Ala-LeuH

(a) Ala-Leu-OMe·HCl: To a solution of 3.0 g (14.83 mmol) of Ala-Leu-OH dissolved in 50 mL of MeOH and cooled to 0ºC is added dropwise 2.43 mL (33.36 mmol) of thionyl chloride. The solution is stirred overnight at room temperature and evaporated to dryness to provide a quantitative yield of the desired product.

(b) Cbz-Gly-Ala-leucine methyl ester: To a solution of 0.414 g (1.98 mmol) of Cbz- Gly-OH and 0.500 g (1.98 mmol) of Ala-Leu-OMe·HCl in CH2Cl2 is added 0.607 mL of TEA, immediately followed by 0.355 mL of DEPC. The solution is stirred overnight and then evaporated. The residue is partitioned between EtOAc and 1 N HCl, the organic layer is washed with saturated NaHCO3 and saturated NaCl, dried (MgSO4) and evaporated to give 650 mg of the crude product.

(c) Moc-Gly-Ala-leucine methyl ester: To a solution of 2.0 g (4.90 mmol) of Cbz-Gly- Ala-leucine methyl ester dissolved in 20 mL of MeOH is added 0.200 g of 10% Pd/C. This mixture is hydrogenated in the presence of 2.45 mL (9.81 mmol) of 4.0 M HCl/dioxane for 2 h, then the reaction is carefully degassed and filtered through celite to remove the catalyst. Evaporation of the MeOH gives 1.45 g of the crude product, which is suspended in 45 mL of CH₂Cl₂ and cooled to 0 °C. To this solution is added 1.45 mL (3.25 mmol) of TEA, followed by 0.362 mL of methyl chloroformate. After stirring overnight, the CH2Cl2 is evaporated and the residue is partitioned between EtOAc and 1 N HCl. The organic phase is separated and washed sequentially with NaHCO3 and NaCl. Drying (MgSO4), evaporation and purification by chromatography gives 0.820 g of the desired product.

(d) Moc-Gly-Ala-leucinol: To a solution of 0.168 g (1.51 mmol) CaCl2 in 25 mL ethanol and 15 mL THF is added 0.250 g of Moc-Gly-Ala-leucine methyl ester. This solution is cooled to -15 °C and 0.114 g (3.02 mmol) of NaBH4 is added in one portion. After stirring for 2 h, the reaction is quenched with 20 mL of 1 N HCl, concentrated on a rotary evaporator and extracted with EtOAc (2x, 50 mL). The combined extracts are washed with saturated NaHCO3 and NaCl, dried (MgSO4) and evaporated. Purification on silica provides 0.167 g of crude product.

(e) Moc-Gly-Ala-LeuH: A solution is prepared by adding 0.418 g (0.989 mmol) of Dess-Martin periodinane to 5 mL of CH2Cl2, followed by stirring for 10 min. Then, 0.100 g (0.330 mmol) of Moc-Gly-Ala-leucinol is added in one portion and the reaction is stirred for 2 h and poured into 25 mL of a solution of saturated NaHCO3 containing 1.72 g (6.93 mmol) of Na2S2O3. After stirring for an additional 10 min, the solution is extracted with EtOAc (3x, 50 mL), dried (MgSO4), and evaporated. Chromatography on silica gave 0.016 g of the desired product.

Synthesis example 4 Synthesis of N-(Methylsulfonyl)-Phe-Gly-Ala-LeuH

(a) N-Ms-Phe-Gly-OH: To a solution of 2.0 g (9.0 mmol) of Phe-Gly-OH dissolved in 9 mL of 1 N NaOH and cooled to 0 °C, 0.766 mL (9.9 mmol) of methanesulfonyl chloride and 9 mL of 1 N NaOH are simultaneously added in separate addition funnels. After the addition is complete, the reaction is stirred at 0 °C for 15 min and at room temperature for 1 h. At this time, the solution is cooled to 0 °C, the pH is adjusted to 9.5, and the solution is washed with EtOAc (1x, 50 mL). The aqueous phase (0°C) is then adjusted to pH = 2.5 (2 N HCl) and washed with EtOAc (3x, 50 mL), dried (MgSO4), filtered and the solvent removed to give 2.0 g of the crude product.

(b) N-Ms-Phe-Gly-Ala-leucinol: A solution is prepared by dissolving 0.500 g (1.67 mmol) of N-Ms-Phe-Gly-OH in 15 mL of THF, cooling to -15ºC and adding 0.366 mL (3.33 mmol) of NMM followed by 0.216 mL (1.67 mmol) of isobutyl chloroformate. This solution is stirred for 5 minutes and 0.374 g (1.67 mmol) of Ala-leucinol·HCl in a mixture of 10 mL of THF and a little DMF is added. Stirring is continued at 0ºC for 15 minutes and 2 h at room temperature. The solution is quenched with 5 mL of 1 N HCl, extracted with EtOAc (3x, 50 mL), the combined extracts are washed with saturated NaHCO3 and saturated NaCl. The resulting organic phase is then dried (MgSO4), filtered, evaporated and chromatographed on silica to give 0.260 g of the desired material.

(c) N-Ms-Phe-Gly-Ala-LeuH: A solution is prepared by adding 0.337 g (0.798 mmol) of Dess-Martin periodinane to 5 mL of CH2Cl2 and stirring for 10 min.

To this solution, 0.125 g (0.266 mmol) of N-Ms-Phe-Gly-Ala-leucinol is added in one portion. The reaction is continued until TLC indicates complete conversion, at which time the solution is poured into 25 mL of saturated NaHCO3 containing 1.8 g (5.586 mmol) of Na2S2O3. After stirring for 10 min, the mixture is extracted with EtOAc (3x, 50 mL). The combined extracts are dried (MgSO4), evaporated, and chromatographed on silica to give 0.048 g of the product.

Synthesis example 5 Synthesis of an aldehyde protease inhibitor

Moc-Leu-OH-L-leucine (5.0 g, 38.2 mmol) is dissolved in 38 mL of 1 N NaOH and cooled to 0 °C. Methyl chloroformate (3.1 mL, 40.0 mmol) is added dropwise while 1 N NaOH is added in a separate addition funnel to maintain the pH at 9.0-9.5. After the addition is complete and the pH is stabilized at 9.0-9.5, the solution is washed with 200 mL of EtOAc, the aqueous phase is then acidified to pH = 2. This mixture is extracted with EtOAc (2x, 100 mL), dried (MgSO4), filtered, and the solvent is removed to give 7.15 g of the crude product.

Moc-Leu-Leucinol: To a solution of 3.5 g (18.52 mmol) of Moc-Leu-OH in 100 mL of THF cooled to -15°C, add 2.04 mL (18.52 mmol) of N-methylmorpholine, immediately followed by 2.4 mL (18.52 mmol) of isobutyl chloroformate. After stirring for 10 min, add 2.37 mL (18.52 mmol) of leucinol in 25 mL of THF and the reaction is stirred for 0.5 h at -15°C and 1 h at room temperature. The mixture is then diluted with 100 mL of H₂O and the THF is evaporated. The remaining aqueous phase is partitioned between EtOAc and 1 N HCl, the organic phase is washed with NaHCO₃. washed, dried (MgSO4) and evaporated to give 5.33 g of the crude product.

Moc-Leu-LeuH: A solution containing 4.4 g (10.41 mmol) of Dess-Martin periodinane suspended in 100 mL of CH2Cl2 is prepared and stirred for 10 min. To this solution is added 1.0 g (3.47 mmol) of Moc-Leu-leucinol and the solution is stirred for 2 h at room temperature and then poured into 100 mL of saturated NaHCO3 containing 18 g (72.87 mmol) of Na2S2O3. This solution is stirred for 10 min and then extracted with EtOAc (2x, 125 mL), dried (MgSO4), and the solvent is evaporated. Chromatography on silica gives 0.550 g of the crude product.

Synthesis example 6

Additional peptide aldehydes are synthesized according to the following procedures. Some of the intermediates are purchased from suppliers and in these cases this is indicated in the procedure. Dess-Martin periodinane is synthesized according to the procedure of Martin, J. Org. Chem. 48 (1983), 4155.

I. Z-Gly-Ala-Leu-OMe: To a solution of Z-Gly-Ala-OH (20.0 g, 0.071 M) and Leu-OMe·HCl (12.9 g, 0.071 M) in 250 mL of dichloromethane is added dropwise 21.9 mL (0.157 M) of triethylamine (TEA) over 10 min. This addition is followed by the addition of 11.9 mL (0.078 M) of diethyl cyanophosphonate (DEPC). The mixture is stirred overnight and the solvent is removed. The residue is dissolved in ethyl acetate and washed with 1 N HCl, saturated NaHCO3 and brine. The solution is dried with MgSO4, filtered and the solvent is removed to give 29.0 g of the product which is homogeneous by TLC. 13C NMR (CDCl3): 15.93; 18.60; 21.77; 22.69; 24.72; 40.80; 44.20; 48.70; 50.87; 52.13; 65.28; 66.84; 127.92; 128.00; 128.41; 136.36; 156.76; 169.31; 172.58; 173.24.

II. Moc-Phe-Gly-Ala-Leu-OMe: Z-Gly-Ala-Leu-OMe (29.0 g, 0.071 M) is dissolved in 300 mL of MeOH and 35 mL of 4.0 HCl in dioxane. To this solvent mixture, 5.8 g of 10% Pd/C is added portionwise. The slurry is degassed with an aspirator and H2 is introduced using a balloon. The slurry is kept under a positive H2 pressure and stirred overnight. The slurry is filtered through celite and a sintered glass funnel and washed thoroughly with MeOH. The solvent is removed and the residue is triturated with ether. The slurry is filtered and the filter cake is dried under reduced pressure. 20.2 g of an off-white powder is obtained. The crude product and Moc-Phe-OH (15.3 g, 0.068 M) are dissolved in 500 mL of CH2Cl2 and 29.9 mL of TEA (0.143 M) are added dropwise, followed by the dropwise addition of 11.7 mL (0.072 M) of DEPC. The mixture is stirred overnight and the solvent is removed. The residue is dissolved in EtOAc and washed with 1 N HCl, saturated NaHCO3 and brine. The organic layer is dried (MgSO4), filtered and the solvent is removed to give 21.3 g of the product.13C NMR (CDCl3): 16.66; 16.83; 20.01; 22.46; 23.41; 25.40; 40.11; 41.72; 43.75; 49.39; 51.37; 52.87; 56.42; 65.92; 77.39; 77.55; 77.81; 78.24; 127.42; 128.96; 129.19; 130.09; 137.41; 157.62; 169.00; 172.63; 173.24; 174.00.

III. Moc-Phe-Gly-Ala-Leucinol: Moc-Phe-Gly-Ala-Leu-OMe (21.3 g, 44.5 mmol) was dissolved in a mixture of 400 mL of EtOH and 250 mL of THF. The solution was cooled to 0 °C and 9.88 g (89.0 mmol) of CaCl2 was added. In 5 min the slurry was homogeneous and 6.73 g (178.0 mmol) of NaBH4 was added portionwise over a period of 5 min. The solution was stirred at 0 °C for 2 h and the reaction was carefully quenched with 1 N HCl. EtOH and THF were removed under reduced pressure and the remaining aqueous mixture was extracted with 500 mL of EtOAc. This organic phase is washed with saturated NaHCO₃, brine and the organic phase is MgSO4. Filtration and removal of solvent gave 20.0 g of off-white crystalline material. Chromatography on silica (3.5% MeOH/CH2Cl2) gave 13.0 g of crude product. Rf = 0.3 (10% MeOH/CH2Cl2); 13C NMR (CDCl3): 17.50; 22.23; 23.12; 24.84; 37.22; 39.76; 43.96; 49.88; 50.93; 52.48; 58.22; 65.27; 98.46; 98.54; 127.04; 128.68; 129.10; 136.62; 157.85; 170.71; 173.85; 174.45.

IV. Moc-Phe-Gly-Ala-Leu-H: 29.9 g (70.7 mmol) of Dess-Martin periodinane was suspended in 500 mL of CH2Cl2 and stirred for 10 min. Moc-Phe-Gly-Ala-leucinol (10.6 g, 23.5 mmol) was dissolved in 100 mL of CH2Cl2 and added at a moderate rate to the periodinane slurry. The mixture was stirred for 1 h and poured into 150 mL of NaHCO3 containing 123 g of Na2S2O3. The mixture was allowed to stir for 15 min and extracted with EtOAc. The organic layer was dried and filtered, then the solvent was removed. Chromatography (3.5% MeOH/CH2Cl2) on silica affords 5.1 g of a pure white solid which is a mixture of the methoxyhemiacetal and the aldehyde. 13C NMR (CDCl3, CD3OD): 17.62; 17.94; 21.53; 21.71; 22.99; 23.30; 23.39; 24.54; 37.05; 37.70; 37.92; 38.24; 42.87; 49.83; 51.79; 52.14; 52.40; 56.75; 57.19; 98.40; 99.18; 127.00; 128.60; 129.06; 136.44; 157.27; 169.19; 169.67; 172.73; 173.40; 200.43.

V. Moc-Phe-OH: L-phenylalanine (5.0 g, 30.2 mmol) is dissolved in 30 mL of 1 N NaOH and cooled to 0 °C. Methyl chloroformate (2.53 mL, 31.8 mmol) is added dropwise while simultaneously adding 30 mL of 1 N NaOH in a separate addition funnel. After the addition is complete, the solution is washed with 200 mL of EtOAc and the aqueous phase is acidified to pH = 2. The mixture is extracted with EtOAc (2x, 100 mL), dried (MgSO4), filtered and the solvent is removed to give 6.0 g of the product. 13C NMR (CDCl3): 37.75; 52.57; 54.64; 128.63; 129.35; 135.74; 156.77; 175.76.

VI. Mac-Phe-OH: To a solution of 1.00 g (2.34 mmol) of Phe-OBn·PTSA in Et₂O at room temperature is added 0.36 mL (2.57 mmol) of TEA. Then 10 mL of MeOH is added, followed by dropwise addition of 0.14 mL of g (2.34 mmol) of methyl isocyanate in 4 mL of Et₂O. The reaction mixture is poured into 50 mL of water and the layers are separated. The organic layer is dried with MgSO₄ and the solvent is removed to give 0.66 g of the product (96% yield). 13C NMR (CDCl₃): 27.05; 38.47; 53.45; 54.64; 65.90; 127.43; 127.85; 128.48; 129.28; 130.27; 135.23; 136.22; 158.17; 173.08. To a solution of the crude product (2.11 mmol) in 25 mL of MeOH is added 0.120 g of Pd/C and the slurry is degassed. The slurry is stirred under a positive H₂ pressure using a balloon for 1.5 h. The slurry is filtered through Celite and the filter cake is with MeOH. The solvent is removed to give 0.430 g of the product. 13C NMR: 26.50; 37.92; 54.28; 126.69; 128.28; 129.28; 136.65; 159.36, 175.33.

VII. Mac-Phe-Gly-Ala-Leucinol: To a solution of 0.200 g of Mac-Phe-OH (0.900 mmol) and 0.253 g of Gly-Ala-Leu-OMe·HCl (0.818 mmol, generated by hydrogenation of I, above, according to the procedure described for compound II) in 15 mL of DMF is added 0.250 mL of TEA (1.80 mmol), followed by the addition of 0.147 mL (0.900 mmol) of DEPC. The mixture is stirred overnight and the solvent is removed. The residue is redissolved in EtOAc and washed sequentially with 0.3 N HCl, saturated NaHCO3, and brine. The solution is dried, filtered, and the solvent is removed to give 0.300 g of the product. The crude product (0.628 mmol) was dissolved in 17 mL of EtOH and cooled to 0 °C. To this solution was added 0.140 g of CaCl2 (1.25 mmol) in 4 mL of THF. To the resulting slurry was added 0.095 g of NaBH4 in one portion. After 45 min, the solution was quenched with water and extracted with EtOAc. The organic phase was dried with MgSO4, filtered and the solvent was removed. Chromatography with 4% MeOH/CH2Cl2 gave 0.200 g of the crude product. 13C NMR (CD3OD): 16.84; 21.05; 22.60; 24.51; 25.66; 37.41; 39.73; 42.67; 49.65; 56.63; 64.33; 126.63; 128.32; 128.96; 137.12; 160.01; 170.45; 173.60; 175.03.

VIII. Mac-Phe-Gly-Ala-Leu-H: To a slurry of Dess-Martin periodinane (0.565 g, 1.33 mmol) in 15 mL of CH2Cl2 is added a suspension of Mac-Phe-Gly-Ala-leucinol (0.200 g, 0.445 mmol) in CH2Cl2 and the resulting slurry is stirred for 0.5 h. The mixture is poured into saturated NaHCO3 containing 2.32% Na2S2O3 and the solution is stirred for 10 min followed by extraction with EtOAc. The organic layer is dried with MgSO4, filtered and the solvent is removed. Chromatograph the residue on silica to give 0.081 g of the product. 13C NMR (10% CD3OD in CDCl3): 17, 18; 17.43; 21.35; 21.55; 23.26; 23.34; 24.40; 24.47; 26.36; 26.60; 37.25; 37, 38; 38.60; 42.86; 42.97; 51.77; 51.93; 54.94; 56.75; 57.00; 98.7; 99.32; 126.87; 128.49; 128.91; 136.51; 159.53; 159.55; 169.93; 170.39; 173.63; 173.85; 174.70.

Cbz = carbobenzyloxy

Gly = Glycine

Ala = Alanine

Leu = Leucin

Phe = phenylalanine

OMe = methyl ester

TEA = Triethylamine

DECP = diethyl cyanophosphonate

TLC = thin layer chromatography

MeOH = methanol

Pd/C = palladium on activated carbon

EtOH = ethanol

THF = Tetrahydrofuran

Mac = Methylaminocarbonyl

Moc = methoxycarbonyl

Etoc = ethoxycarbonyl

Ms = methanesulfonyl

Proteolytic enzyme

Another essential ingredient in the present liquid detergent compositions is an active proteolytic enzyme. Mixtures of proteolytic enzymes are also included. The proteolytic enzyme can be derived from animals, plants or microorganisms (preferred). The proteases for use in the detergent compositions herein include (but are not limited to) trypsin, subtilisin, chymotrypsin and elastase type proteases. Preferred for use herein are subtilisin type proteolytic enzymes. Particularly preferred is a bacterial serine proteolytic enzyme obtained from Bacillus subtilis and/or Bacillus licheniformis. Protease enzymes are typically present in such liquid detergent compositions in sufficient amounts to provide 0.005 to 0.1 Anson Units (AU) of activity per gram of composition.

Suitable proteolytic enzymes include Alcalase® (preferred), Esperase® and Savinase® from Novo Industri A/S (Copenhagen, Denmark); Maxatase®, Maxacal® and Maxapem 15® (protein-modified Maxacal®) from Gist-Brocades (Delft, The Netherlands); and Subtilisin BPN and BPN' (preferred), which are commercially available. Preferred proteolytic enzymes are also modified bacterial serine proteases, such as those produced by Genencor International, Inc. (San Francisco, California), described in European Patent 251,446, filed April 28, 1987 (especially pages 17, 24 and 98), referred to herein as "Protease B", and in U.S. Patent 5,030,378, Venegas, issued July 9, 1991, which refers to a modified bacterial serine proteolytic enzyme (Genencor International), referred to herein as "Protease A" (corresponds to BPN'). See especially columns 2 and 3 of U.S. Patent 5,030,378 for a complete description, including amino acid sequence, of Protease A and variants thereof. Preferred proteolytic enzymes are thus selected from the group consisting of Alcalase® (Novo Industri A/S), BPN', Protease A and Protease B (Genencor), and mixtures thereof. Protease B is most preferred.

Another preferred protease, referred to as "Protease D", is a carbonyl hydrolase variant having an amino acid sequence not found in nature, which is derived from a precursor carbonyl hydrolase by substituting a plurality of amino acid residues with another amino acid at a position in the carbonyl hydrolase corresponding to position +76, preferably also in combination with one or more amino acid residue positions corresponding to those selected from the group consisting of +99, +101, +103, +104, +107, +123, +27, +105, +109, +126, +128, +135, +156, +166, +195, +197, +204, +206, +210, +216, +217, +218, +222, +260, +265 and/or +274, according to the numbering of Bacillus amyloliquefaciens subtilisin as described in WO 95/10615, published April 20, 1995 by Genencor International.

Useful proteases are also described in PCT publications: WO 95/30010, published November 9, 1995 by The Procter & Gamble Company; WO 95/30011, published November 9, 1995 by The Procter & Gamble Company; and WO 95/29979, published November 9, 1995 by The Procter & Gamble Company.

Calcium

Any water-soluble calcium salt may be used as the source of calcium ions, including calcium acetate, calcium formate, calcium xylene sulfonate, and calcium propionate. Divalent ions such as zinc and magnesium ions may partially replace the calcium ions. Thus, in the liquid detergent compositions herein, the source of calcium ions may be partially replaced by a source of other divalent ions.

The calcium useful herein is accessible to the enzyme. Therefore, the claimed compositions are substantially free of sequestrants, for example polyacids, which are capable of forming calcium complexes which are soluble in the composition. However, small amounts of sequestrants such as polyacids or mixtures of polyacids may be used. The calcium accessible to the enzyme is defined as the amount of calcium ions which are effectively accessible to the enzyme component. From a practical standpoint, the calcium accessible to the enzyme is therefore the soluble calcium in the composition in the absence of any storage sequestrants, e.g. having a complexation equilibrium constant with calcium equal to or greater than 1.5 at 20°C.

Boric acid

The compositions herein optionally contain from about 0.25 to about 10% by weight, preferably from about 0.5 to about 5% by weight, more preferably from about 0.75 to about 3% by weight, of boric acid or a compound capable of forming boric acid in the composition (calculated on a boric acid basis). Boric acid is preferred, although other compounds such as boron oxide, borax and other alkali metal borates (e.g. sodium ortho-, meta-, pyroborate or sodium pentaborate) are suitable. Substituted boric acids (e.g. phenylboric acid, butaneboric acid and p-bromophenylboric acid) may also be used instead of boric acid.

The compositions of the invention may also contain polyols, especially diols containing only carbon, hydrogen and oxygen atoms. They preferably contain from about 2 to about 6 hydroxy groups. Examples include propylene glycol (especially 1,2-propanediol, which is preferred), ethylene glycol, glycerol, sorbitol, mannitol, glucose and mixtures thereof. The polyol generally comprises from about 1 to about 15%, preferably from about 1.5 to about 10%, more preferably from about 2 to about 7%, by weight of the composition.

Detergent

An effective amount of from 1 to 95% by weight, preferably from 8 to 70% by weight, of detergent surfactant is another essential ingredient of the present invention. The detergent surfactant may be selected from the group consisting of anionic, nonionic, cationic, ampholytic and zwitterionic surfactants, and mixtures thereof. By selecting the type and amount of detergent surfactant together with other adjunct ingredients disclosed herein, the present detergent compositions can be formulated for use in connection with laundry cleaning or in other various cleaning applications, especially including dishwashing. The particular surfactants used can therefore vary widely depending on the particular use contemplated.

The advantages of the present invention are particularly evident in compositions containing ingredients that are aggressive to enzymes, such as certain detergent builders and surfactants. These include (but are not limited to) anionic surfactants such as an alkyl ether sulfate, linear alkyl benzene sulfonate, alkyl sulfate, etc. Suitable surfactants are described below.

Anionic surfactants

One type of anionic surfactant which may be used includes alkyl ester sulfonates. These are desirable because they can be prepared from renewable, non-petrochemical feedstocks. Preparation of the alkyl ester sulfonate surfactant component can be carried out according to known methods disclosed in the technical literature. For example, linear esters of C8-C20 carboxylic acids can be sulfonated with gaseous SO3 according to "The Journal of the American Oil Chemists Society", 52 (1975), pp. 323-329. Suitable starting materials would include natural fatty substances such as those derived from tallow, palm and coconut oils, etc.

The preferred alkyl ester sulfonate surfactant, especially for washing applications, includes alkyl ester sulfonate surfactants having the structural formula:

wherein R3 is C8-C20 hydrocarbyl, preferably alkyl or a combination thereof; R4 is C1-C6 hydrocarbyl, preferably alkyl or a combination thereof; and M is a cation which forms a soluble salt. Suitable salts include metal salts such as sodium, potassium and lithium salts, and substituted or unsubstituted ammonium salts such as methyl, dimethyl, trimethyl and quaternary ammonium cations, e.g. tetramethylammonium and dimethylpiperidinium, and cations derived from alkanolamines, e.g. monoethanolamine, diethanolamine and triethanolamine. Preferably R3 is C10-C16 alkyl and R4 is methyl, ethyl or isopropyl. Particularly preferred are the methyl ester sulfonates in which R³ is C₁₄-C₁₆ alkyl.

Alkyl sulfate surfactants are another important type of anionic surfactant for use herein. In addition to providing excellent overall cleaning ability when used in combination with polyhydroxy fatty acid amides (see below), including good grease/oil cleaning over a wide range of temperatures, wash concentrations and wash times, the solution of alkyl sulfates as well as improved formability in liquid detergent formulations can be obtained. They are water-soluble salts or acids of the formula ROSO₃M, wherein R is preferably a C₁₀-C₂₄ hydrocarbyl, preferably an alkyl or hydroxyalkyl having a C₁₀-C₂₀ alkyl moiety, more preferably a C₁₂-C₁₈ alkyl or hydroxyalkyl, and M is H or a cation, e.g. B. an alkali metal cation (e.g., sodium, potassium, lithium), substituted or unsubstituted ammonium cations such as methyl, dimethyl and trimethyl ammonium, and quaternary ammonium cations such as tetramethyl ammonium and dimethylpiperidinium, as well as cations derived from alkanolamines such as ethanolamine, diethanolamine, triethanolamine and mixtures thereof, and the like. Typically, alkyl chains of C₁₂-C₁₆ are preferred for low wash temperatures (e.g., below about 50°C), and C₁₆-C₁₈ alkyl chains are preferred for higher wash temperatures (e.g., above about 50°C).

Alkoxylated alkyl sulfate surfactants are another category of useful anionic surfactant. These surfactants are water-soluble salts or acids, typically having the formula RO(A)mSO3M, wherein R is an unsubstituted C10-C24 alkyl or hydroxyalkyl group having a C10-C24 alkyl moiety, preferably a C12-C20 alkyl or hydroxyalkyl, more preferably a C12-C18 alkyl or hydroxyalkyl, A is an ethoxy or propoxy group, m is greater than zero, usually between about 0.5 and about 6, more preferably between about 0.5 and about 3, and MH is or a cation which may be, for example, a metal cation (e.g., sodium, potassium, lithium, calcium, magnesium, etc.), ammonium or a substituted ammonium cation. Ethoxylated alkyl sulfates and propoxylated alkyl sulfates are included herein. Specific examples of substituted ammonium cations include methyl, dimethyl, trimethylammonium and quaternary ammonium cations such as tetramethylammonium, dimethylpiperidinium, and cations derived from alkanolamines, e.g. monoethanolamine, diethanolamine and triethanolamine, and mixtures thereof. Exemplary surfactants are C₁₂-C₁₈ alkyl polyethoxylate (1,0) sulfate, C₁₂-C₁₈ alkyl polyethoxylate (2,25) sulfate, C₁₂-C₁₈ alkyl polyethoxylate (3,0) sulfate and C₁₂-C₁₈ alkyl polyethoxylate (4,0) sulfate, wherein M is suitably selected from sodium and potassium.

Other anionic surfactants

Other anionic surfactants useful for cleaning purposes may also be included in the compositions herein. These may be salts (including, for example, sodium, potassium, ammonium and substituted ammonium salts such as mono-, di- and triethanolamine salts) of soap, linear C9-C20 alkylbenzene sulfonates, primary or secondary C8-C22 alkane sulfonates, C8-C24 olefin sulfonates, sulfonated polycarboxylic acids prepared by sulfonation of the pyrolyzed product of alkaline earth metal citrates, e.g. B. as described in British Patent Specification No. 1,082,179, alkylglycerol sulfonates, fatty acylglycerol sulfonates, fatty oleylglycerol sulfates, alkylphenol ethylene oxide ether sulfates, paraffin sulfonates, alkyl phosphates, isothionates such as the acyl isothionates, N-acyl taurates, fatty acid amides of methyl tauride, alkyl succinamates and sulfosuccinates, monoesters of sulfosuccinate (particularly saturated and unsaturated C₁₂-C₁₈ monoesters), diesters of sulfosuccinate (particularly saturated and unsaturated C₆-C₁₄ diesters), N-acyl sarcosinates, sulfates of alkylpolysaccharides such as the sulfates of alkylpolyglucoside (the nonionic nonsulfated compounds described below), branched primary alkyl sulfates, alkyl polyethoxycarboxylates such as those of the formula RO(CH₂CH₂O)kCH₂COO⁻M⁺, where R is C₈-C₂₂ alkyl, k is an integer from 0 to 10, and M is a soluble salt forming cation, and fatty acids esterified with isethionic acid and neutralized with sodium hydroxide. Resin acids and hydrogenated resin acids are also suitable, such as rosin or hydrogenated rosin, as well as resin acids and hydrogenated resin acids present in or derived from tall oil. Further examples are given in "Surface Active Agents and Detergents" (Vols. I and II by Schwartz, Perry and Berch). A variety of such surfactants are also generally disclosed in US Patent 3,929,678, issued December 30, 1975 to Laughlin et al., at column 23, line 58 through column 29, line 23.

Non-ionic detergent surfactants

Suitable nonionic detersive surfactants are generally disclosed in U.S. Patent 3,929,678, Laughlin et al., issued December 30, 1975, at column 13, line 14 through column 16, line 6. Exemplary nonlimiting classes of useful nonionic surfactants are listed below.

The polyethylene, polypropylene and polybutylene oxide condensates of alkylphenols. Generally, the polyethylene oxide condensates are preferred. These compounds include the condensation products of alkylphenols having an alkyl group containing from about 6 to about 12 carbon atoms in either a straight chain or branched chain configuration with the alkylene oxide. In a preferred embodiment, the ethylene oxide is present in an amount corresponding to about 5 to about 25 moles ethylene oxide per mole of alkylphenol. Commercially available nonionic surfactants of this type include Igepal® CO-630, sold by GAF Corporation; and Triton® X-45, X-114, X-100 and X-102, all sold by Rohm & Haas Company. These compounds are commonly referred to as alkylphenol alkoxylates (e.g., alkylphenol ethoxylates).

The condensation products of aliphatic alcohols with about 1 to about 25 moles of ethylene oxide. The alkyl chain of the aliphatic alcohol can be either straight or branched, primary or secondary, and generally contains from about 8 to about 22 carbon atoms. Particularly preferred are the condensation products of alcohols having an alkyl group containing from about 10 to about 20 carbon atoms with about 2 to about 18 moles of ethylene oxide per mole of alcohol. Examples of commercially available nonionic surfactants of this type include Tergitol® 15-S-9 (the condensation product of a linear C11-C15 secondary alcohol with 9 moles of ethylene oxide), Tergitol® 24-L-6-NMW (the condensation product of a primary C12-C14 alcohol with 6 moles of ethylene oxide having a narrow molecular weight distribution), both sold by Union Carbide Corporation; Neodol® 45-9 (the condensation product of a linear C₁₄-C₁₅ alcohol with 9 moles of ethylene oxide), Neodol® 23-6.5 (the condensation product of a linear C₁₂-C₁₃ alcohol with 6.5 moles of ethylene oxide), Neodol® 45-7 (the condensation product of a linear C₁₄-C₁₅ alcohol with 7 moles of ethylene oxide) and Neodol® 45-4 (the condensation product of a linear C₁₄-C₁₅ alcohol with 4 moles of ethylene oxide), sold by Shell Chemical Company, and Kyro® EOB (the condensation product a C₁₃-C₁₅ alcohol with 9 moles of ethylene oxide), distributed by The Procter & Gamble Company. This category of nonionic surfactant is commonly referred to as "alkyl ethoxylates".

The condensation products of ethylene oxide with a hydrophobic base, which are formed by the condensation of propylene oxide with propylene glycol. The hydrophobic portion of these compounds preferably has a molecular weight of about 1,500 to about 1,800 and exhibits insolubility in water. The addition of polyoxyethylene groups to this hydrophobic portion tends to increase the water solubility of the molecule as a whole, and the liquid character of the product is maintained up to the point where the polyoxyethylene content is about 50% of the total weight of the condensation product, which corresponds to condensation with up to about 40 moles of ethylene oxide. Examples of compounds of this type include certain of the commercially available Pluronic® surfactants sold by BASF.

The condensation products of ethylene oxide with the product resulting from the reaction of propylene oxide and ethylenediamine. The hydrophobic group of these products consists of the reaction product of ethylenediamine and excess propylene oxide and generally has a molecular weight of about 2,500 to about 3,000. This hydrophobic group is condensed with ethylene oxide to the extent that the condensation product contains about 40 to about 80 weight percent polyoxyethylene and has a molecular weight of about 5,000 to about 11,000. Examples of this type of nonionic surfactant include certain of the commercially available Tetronic® compounds sold by BASF.

Semipolar nonionic surfactants are a special category of nonionic surfactants which include water-soluble amine oxides containing one alkyl group of about 10 to about 18 carbon atoms and two groups selected from the group consisting of alkyl groups and hydroxyalkyl groups having about 1 to about 3 carbon atoms; water-soluble phosphine oxides containing one alkyl group of about 10 to about 18 carbon atoms and two groups selected from the group consisting of alkyl groups and hydroxyalkyl groups having about 1 to about 3 carbon atoms; and water-soluble sulfoxides containing one alkyl group of about 10 to about 18 carbon atoms and one group selected from the group consisting of alkyl and hydroxyalkyl groups having about 1 to about 3 carbon atoms.

Semipolar nonionic detergent surfactants include the amine oxide surfactants with the formula:

wherein R3 is an alkyl, hydroxyalkyl or alkylphenyl group or mixtures thereof having from about 8 to about 22 carbon atoms; R4 is an alkylene or hydroxyalkylene group having from about 2 to about 3 carbon atoms or mixtures thereof; x is 0 to about 3; and each R5 is an alkyl or hydroxyalkyl group having from about 1 to about 3 carbon atoms or a polyethylene oxide group having from about 1 to about 3 ethylene oxide groups. The R5 groups may be linked together to form a ring structure, for example, via an oxygen or nitrogen atom.

These amine oxide surfactants include, in particular, C₁₀-C₁₈ alkyldimethylamine oxides and C₈-C₁₂ alkoxyethyldihydroxyethylamine oxides.

Alkyl polysaccharides, disclosed in U.S. Patent 4,565,647, Llenado, issued January 21, 1986, which have a hydrophobic group of about 6 to about 30 carbon atoms, preferably about 10 to about 16 carbon atoms, and a hydrophilic polysaccharide group, e.g., a polyglycoside, of about 1.3 to about 10, preferably about 1.3 to about 3, most preferably about 1.3 to about 2.7 saccharide groups. Any reducing saccharide having 5 or 6 carbon atoms may be used, e.g., the glucosyl groups may be replaced by glucose, galactose, and galactosyl groups. (Optionally, the hydrophobic group is attached in the 2-, 3-, 4-, etc. positions, thereby providing a glucose or galactose as opposed to a glucoside or galactoside). The intersaccharide bonds can be, for example, between the 1-position of the additional saccharide groups and the 2-, 3-, 4- and/or 6-positions of the preceding saccharide groups.

Optionally, and less preferably, a polyalkylene oxide chain may be present connecting the hydrophobic group and the polysaccharide group. The preferred alkylene oxide is ethylene oxide. Typical hydrophobic groups include alkyl groups, either saturated or unsaturated, branched or unbranched, containing from about 8 to about 18, preferably from about 10 to about 16, carbon atoms. Preferably, the alkyl group is a straight chain saturated alkyl group. The alkyl group may contain up to about 3 hydroxy groups and/or the polyalkylene oxide chain may contain up to about 10, preferably less than 5, alkylene oxide groups. Suitable alkyl polysaccharides are octyl, nonyl, decyl, undecyldodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl and octadecyl di-, tri-, tetra-, penta- and hexaglucosides, galactosides, lactosides, glucoses, fructosides, fructoses and/or galactoses. Suitable mixtures include coconut alkyl di-, tri-, tetra and pentaglucosides and tallow alkyl tetra-, penta- and hexaglucosides.

The preferred alkyl polyglycosides have the formula:

R²O(CnH2nO)t(Glycosyl)x

wherein R² is selected from the group consisting of alkyl, alkylphenyl, hydroxyalkyl, hydroxyalkylphenyl, and mixtures thereof, wherein the alkyl groups contain from about 10 to about 18, preferably from about 12 to about 14 carbon atoms; n is 2 or 3, preferably 2; t is 0 to about 10, preferably 0; and x is from about 1.3 to about 10, preferably from about 1.3 to about 3, most preferably from about 1.3 to about 2.7. The glycosyl is preferably derived from glucose. To prepare these compounds, the alcohol or alkylpolyethoxy alcohol is first formed and then reacted with glucose or a source of glucose to form the glucoside (bonding in the 1-position). The additional glycosyl groups can then be bonded between their 1-position and the previous glycosyl groups in the 2-, 3-, 4- and/or 6-position, preferably predominantly in the 2-position.

Fatty acid amide surfactants with the formula:

wherein R6 is an alkyl group having from about 7 to about 21 (preferably from about 9 to about 17) carbon atoms and each R7 is selected from the group consisting of hydrogen, C1-C4 alkyl, C1-C4 hydroxyalkyl and -(C2H4O)xH, where x varies from about 1 to about 3.

Preferred amides are C8-C20 ammonia amides, monoethanol amides, diethanol amides and isopropanol amides.

Cationic surfactants

Cationic detersive surfactants may also be included in the detergent compositions of the invention. Cationic surfactants include the ammonium surfactants such as alkyldimethylammonium halides and those surfactants having the formula:

[R²(OR³y][R⁴(OR³)y]₂R⁵N⁺X⁻

wherein R2 is an alkyl or alkylbenzyl group having from about 8 to about 18 carbon atoms in the alkyl chain; each R3 is selected from the group consisting of -CH2CH2-, -CH2CH(CH3)-, -CH2CH(CH2OH)-, CH2CH2CH2, and mixtures thereof; each R4 is selected from the group consisting of C1-C4 alkyl, C1-C4 hydroxyalkyl, benzyl, ring structures formed by linking the two R4 groups, -CH2CHOH-CHOHCO- R6CHOHCH2OH, wherein R6 is is any hexose or hexose polymer having a molecular weight of less than about 1,000, and hydrogen when y is not 0; R5 has the same meaning as R4 or is an alkyl chain wherein the total number of carbon atoms of R2 plus R5 is not more than about 18; each y is 0 to about 10 and the sum of the y values is 0 to about 15; and X is any compatible anion.

Other cationic surfactants useful herein are also described in U.S. Patent 4,228,044, Cambre, issued October 14, 1980.

Other surfactants

Ampholytic surfactants can be incorporated into the detergent compositions herein. These surfactants can be generally described as aliphatic derivatives of secondary or tertiary amines or aliphatic derivatives of heterocyclic secondary and tertiary amines, wherein the aliphatic moiety can be straight chain or branched. One of the aliphatic substituents contains at least about 8 carbon atoms, typically about 8 to about 18 carbon atoms, and at least one contains an anionic water solubilizing group, e.g., carboxy, sulfonate, or sulfate. See U.S. Patent No. 3,929,678 to Laughlin et al., issued December 30, 1975, at column 19, lines 18-35, for examples of ampholytic surfactants.

Zwitterionic surfactants can also be incorporated into the detergent compositions herein. These surfactants can be generally described as derivatives of secondary and tertiary amines, derivatives of heterocyclic secondary and tertiary amines, or derivatives of quaternary ammonium, quaternary phosphonium or tertiary sulfonium compounds. See U.S. Patent No. 3,929,678 to Laughlin et al., issued December 30, 1975, at column 19, line 18 through column 22, line 48, for examples of zwitterionic surfactants. Ampholytic and zwitterionic surfactants are generally used in combination with one or more anionic and/or nonionic surfactants.

Polyhydroxy fatty acid amide surfactant

The liquid detergent compositions herein may also contain an enzyme enhancing amount of polyhydroxy fatty acid amide surfactant. By "enzyme enhancing amount" it is meant that the formulator of the composition can select an amount of polyhydroxy fatty acid amide to be incorporated into the compositions which will improve the enzyme cleaning performance of the detergent composition. For conventional enzyme levels, the incorporation of about 1% by weight of polyhydroxy fatty acid amide generally improves enzyme performance.

The detergent compositions herein typically comprise about 1% by weight of polyhydroxy fatty acid amide surfactant, preferably about 3 to about 30% of the polyhydroxy fatty acid amide. The polyhydroxy fatty acid amide surfactant component comprises compounds having the structural formula:

wherein R¹ is H, C₁-C₄ hydrocarbyl, 2-hydroxyethyl, 2-hydroxypropyl, or a mixture of these, preferably C₁-C₄ alkyl, more preferably C₁ or C₂ alkyl, most preferably C₁ alkyl (i.e., methyl); and R² is C₅-C₃₁ hydrocarbyl, preferably straight chain C₇-C₁₇ alkyl or alkenyl, more preferably straight chain C₇-C₁₇ alkyl or alkenyl, most preferably straight chain C₁₁-C₁₅ alkyl or alkenyl, or mixtures thereof; and Z is a polyhydroxyhydrocarbyl having a linear hydrocarbyl chain with at least 3 hydroxyl groups directly attached to the chain, or an alkoxylated derivative (preferably ethoxylated or propoxylated) thereof. Z is preferably derived from a reducing sugar in a reductive amination reaction; more preferably Z is a glycityl. Suitable reducing sugars include glucose, fructose, maltose, lactose, galactose, mannose and xylose. As starting materials, high dextrose corn syrup, high fructose corn syrup and high maltose corn syrup as well as the individual sugars listed above can be used. These corn syrups can provide a mixture of sugar components for Z. It should be understood that other suitable starting materials are not excluded. Z is preferably selected from the group consisting of -CH₂-(CHOH)n-CH₂OH, -CH(CH₂OH)-(CHOH)n-1-CH₂OH, -CH₂-(CHOH)₂-(CHOR')(CHOH)-CH₂OH and alkoxylated derivatives thereof, wherein n is an integer from 3 to 5 inclusive and R' is H or a cyclic or aliphatic monosaccharide. Most preferred are glycityls wherein n is 4, especially -CH₂-(CHOH)₄-CH₂OH.

R' can be, for example, N-methyl, N-ethyl, N-propyl, N-isopropyl, N-butyl, N-2-hydroxyethyl or N-2-hydroxypropyl.

R²-CO-N< can be, for example, cocamide, stearamide, oleamide, lauramide, myristamide, capricamide, palmitamide, tallowamide, etc.

Z can be 1-deoxyglucityl, 1-deoxyfructityl, 1-deoxymaltityl, 1-deoxylactityl, 1-deoxygalactityl, 1-deoxymannityl, 1-deoxymaltotriotityl etc.

Methods for preparing polyhydroxy fatty acid amides are known in the art. In general, they can be prepared by reacting an alkylamine with a reducing sugar in a reductive amination reaction to form a corresponding N-alkylpolyhydroxyamine and then reacting the N-alkylpolyhydroxyamine with an aliphatic fatty ester or a triglyceride in a condensation/amination step to form the N-alkyl-N-polyhydroxy fatty acid amide product. Methods for preparing compositions containing polyhydroxy fatty acid amides are described, for example, in GB Patent Specification 809,060, published February 18, 1959 by Thomas Hedley & Co., Ltd.; in US Patent 2,965,576, issued December 20, 1960 to Wilson E. R.; and U.S. Patent 2,703,798, Schwartz Anthony M., issued March 8, 1955; and U.S. Patent 1,985,424, issued December 25, 1934 to Piggott.

Second enzyme

Preferred compositions herein further comprise a performance enhancing amount of a detergent compatible second enzyme. By "detergent compatible" is meant compatibility with the other ingredients of a liquid detergent composition such as detersive surfactant and detergent builder. These second enzymes are preferably selected from the group consisting of lipase, amylase, cellulase and mixtures thereof. The term "second enzyme" excludes the proteolytic enzymes discussed above, so that any composition comprising a second enzyme contains at least two types of enzyme, including at least one proteolytic enzyme. The amount of second enzyme used in the composition varies according to the enzyme type. Generally, preferably about 0.0001 to 0.3% by weight, more preferably 0.001 to 0.1% by weight, of these second enzymes are used. Mixtures of the same class of enzymes (e.g. lipase) or of two or more classes (e.g. cellulase and lipase) may be used. Purified or unpurified forms of the enzyme may be used.

Any lipolytic enzyme suitable for use in a liquid detergent composition can be used in these compositions. Suitable lipase enzymes for use herein include those derived from bacteria and fungi.

Suitable bacterial lipases include those produced by microorganisms of the Pseudomonas group, e.g. Pseudomonas stutzeri ATCC 19.154 as disclosed in British Patent 1,372,034. Suitable lipases include those which show a positive immunological cross-reaction with the antibody to the lipase produced by the microorganism Pseudomonas fluorescens IAM 1057. This lipase and a process for its purification are described in Japanese Patent Application 53-20487, which was published on February 24, 1978. This lipase is available from Amano Pharmaceutical Co., Ltd., Nagoya, Japan, under the trade name Lipase P "Amano", hereinafter referred to as "Amano-P®". Such lipases should show a positive immunological cross-reaction with the Amano-P antibody using the conventional and well-known immunodiffusion method according to Ouchterlony (Acta. Med. Scan. 133 (1950), pp. 76-79). These lipases and a method for their immunological cross-reaction with Amano-P are also described in U.S. Patent 4,707,291, Thom et al., issued November 17, 1987. Typical examples of these are Amano-P lipase, the lipase from Pseudomonas fragi FERM P-1339 (available under the trade name Amano-B), the lipase from Pseudomonas nitroreducens Var. lipolyticum FERM P-1338 (available under the trade name Amano-CES), lipases from Chromobacter viscosum, e.g. Chromobacter viscosum Var. lipolyticum NRRLB, commercially available from Toyo Jozo Co., Tagata, Japan, and further Chromobacter viscosum lipases from US Biochemical Corp., USA, and Disoynth Co., Netherlands, and lipases from Pseudomonas gladioli.

Suitable fungal lipases include those that can be produced by Humicola lanuginosa and Thermomyces lanuginosus. Most preferred is the lipase obtained by cloning the gene from Humicola lanuginosa and expressing the gene in Aspergillus oryzae as described in European Patent Application 0 258 068 (Novo Industri A/S), which is commercially available from Novo Nordisk A/S under the trade name Lipolase®.

In these compositions, about 10 to 18,000, preferably about 60 to 6,000, lipase units per gram (LU/g) of lipase can be used. One lipase unit is the amount of lipase which produces 1 mmol of titratable butyric acid per minute at a stable pH, where the pH is 9.0, the temperature is 30°C, the substrate is an emulsion of 3.3% by weight olive oil and 3.3% gum arabic, in the presence of 13 mmol/L C++ and 20 mmol/L NaCl in 5 mmol/Tris buffer.

Any cellulase suitable for use in a liquid detergent composition can be used in these compositions. Suitable cellulase enzymes for use herein include those derived from bacteria and fungi. Preferably they have a pH optimum between 5 and 9.5. About 0.0001 to 0.1% by weight of cellulase can be used.

Suitable cellulases are disclosed in US Patent 4,435,307, Barbesgaard et al., issued March 6, 1984, incorporated herein by reference, which discloses a fungal cellulase produced by Humicola insolens. Suitable cellulases are also disclosed in GB-A-2,075,028; GB-A-2,095,275 and DE-OS-2,247,832.

Examples of such cellulases are cellulases produced by a strain of Humicola insolens (Humicola grisea Var. thermoidea), particularly the Humicola strain DSM 1800, and cellulases produced by a fungus of Bacillus N or a cellulase-212 producing fungus of the genus Aeromonas, and a cellulase extracted from the midgut gland of a marine mollusk (Dolabella auricula Solander).

Any amylase suitable for use in a liquid detergent composition can be used in these compositions. Amylases include, for example, amylases obtained from a particular strain of B. licheniformis, which are more fully described in British Patent Specification No. 1,296,839 (Novo). Amylolytic proteins include, for example, Rapidase®, International Bio-Synthetics, Inc., and Termamyl®, Novo Industries.

About 0.0001 to 0.55% by weight, preferably 0.0005 to 0.1% by weight, amylase may be used.

Optional components

Detergent builders may optionally be included in the compositions herein, particularly for laundry detergent compositions. Inorganic as well as organic builders may be used. When present, the compositions typically comprise at least about 1% of a builder, which may be either an inorganic or organic builder. Liquid laundry detergent formulations preferably comprise about 3 to 30% by weight, more preferably about 5 to 20% by weight, of a detergent builder.

Inorganic detergent builders include, but are not limited to, the alkali metal, ammonium and alkanolammonium salts of polyphosphates (exemplified by the tripolyphosphates, pyrophosphates and the glassy polymeric metaphosphates), phosphonates, phytic acid, silicates, carbonates (including bicarbonates and sesquicarbonates), sulfates and aluminosilicates. Borate builders and builders containing borate-forming materials which can generate borate during detergent storage or under wash conditions (hereinafter collectively referred to as "borate builders") may also be used. Preferably, non-borate builders intended for use under wash conditions of less than about 50°C, especially less than about 40°C, are used in the compositions of the invention.

Examples of silicate builders are the alkali metal silicates, particularly those having a SiO2:Na2O ratio in the range of 1.6:1 to 3.2:1, and layered silicates such as the sodium layered silicates described in U.S. Patent 4,664,839 issued May 12, 1987 to Rieck H. P. However, other silicates may also be useful, such as magnesium silicate, which can serve as a solidifying agent in granular formulations, as a stabilizer for oxygen bleaches, and as a component of foam control systems.

Examples of carbonate builders are the alkaline earth and alkali metal carbonates, including sodium carbonate and sesquicarbonate, and mixtures thereof with ultrafine Calcium carbonate as disclosed in German Patent Application No. 2,321,001, published on November 15, 1973.

Aluminosilicate builders are useful in the present invention. Aluminosilicate builders are very important in most currently marketed granular heavy-duty detergent compositions and can also be a significant builder ingredient in liquid detergent formulations. Aluminosilicate builders include those having the empirical formula:

Mz(zAlO₂·ySiO₂)

wherein M is sodium, potassium, ammonium or substituted ammonium; z is about 0.5 to about 2; and y is 1; this material has a magnesium ion exchange capacity of at least about 50 milligram equivalents of CaCO₃ hardness per gram of the anhydrous aluminosilicate. Preferred aluminosilicates are zeolite builders having the formula:

Na 2 [(AlO 2 )z(SiO 2 )y] xH 2 O

wherein z and y are integers of at least 6, the molar ratio of z to y is in the range of 1.0 to about 0.5, and x is an integer of about 15 to about 264.

Useful aluminosilicate ion exchange materials are commercially available. These aluminosilicates may have a crystalline or amorphous structure and may be naturally occurring aluminosilicates or synthetically derived. A process for preparing aluminosilicate ion exchange materials is disclosed in U.S. Patent 3,985,669, Krummel et al., issued October 12, 1976. Preferred synthetic crystalline aluminosilicate ion exchange materials useful herein are available under the designations Zeolite A, Zeolite P (B) and Zeolite X. In a particularly preferred embodiment, the crystalline aluminosilicate ion exchange material has the formula:

Na 12 [(AlO 2 ) 12 (SiO 2 )n] xH 2 O

where x is from about 20 to about 30, especially about 27. This material is known as zeolite A. Preferably, the aluminosilicate has a particle size of from about 0.1 to 10 microns in diameter.

Specific examples of polyphosphates are the alkali metal tripolyphosphates, sodium, potassium and ammonium pyrophosphate, sodium and potassium and ammonium pyrophosphate, sodium and potassium orthophosphate, sodium polymetaphosphate, in which the degree of polymerization is in the range of about 6 to about 21, and salts of phytic acid.

Examples of phosphonate builder salts are the water-soluble salts of ethane-1-hydroxy-1,1-diphosphonate, especially the sodium and potassium salts; the water-soluble salts of methylenediphosphonic acid, e.g. the trisodium and tripotassium salts; and the water-soluble salts of substituted methylenediphosphonic acids, such as the trisodium and tripotassium methylidene, isopyropylidene, benzylmethylidene and halomethylidene phosphonates. Phosphonate builder salts of the types mentioned above are disclosed in U.S. Patent Nos. 3,159,581 and 3,213,030, respectively, issued December 1, 1964 and October 19, 1965 to Diehl; U.S. Patent No. 3,422,021, issued January 14, 1969 to Roy; and U.S. Patent Nos. 3,400,148 and 3,422,137, issued September 3, 1968 and January 14, 1969 to Quimby.

Organic detergent builders preferred for the purposes of the present invention include a wide variety of polycarboxylate compounds. As used herein, "polycarboxylate" refers to compounds having a plurality of carboxylate groups, preferably at least 3 carboxylate groups.

The polycarboxylate builder can generally be added to the composition in the acid form, but can also be added in the form of a neutralized salt. If the salt form is used, alkali metals such as sodium, potassium and lithium, or alkanolammonium salts are preferred.

The polycarboxylate builders include a variety of categories of useful materials. One important category of polycarboxylate builders comprises the ether polycarboxylates. A number of ether polycarboxylates have been disclosed for use as detergent builders. Examples of useful ether polycarboxylates include oxydisuccinate as disclosed in Berg, U.S. Patent 3,128,287, issued April 7, 1964, and Lamberti et al., U.S. Patent 3,635,830, issued January 18, 1972.

A particular type of ether polycarboxylates useful as builders in the present invention also includes those having the general formula:

CH(A)(COOX)-CH(COOX)-O-CH(COOX)-CH(COOX)(B)

wherein A is H or OH; B is H or -O-CH(COOX)-CH2(COOX); and X is H or a salt-forming cation. For example, if in the above general formula, A and B are both H, then the compound is oxydisuccinic acid and its water-soluble salts. If A is OH and B is H, then the compound is tartrate monosuccinic acid (TMS) and its water-soluble salts. If A is H and B is -O-CH(COOX)-CH2(COOX), then the compound is tartrate disuccinic acid (TDS) and its water-soluble salts. Mixtures of these builders are particularly preferred for use herein. Particularly preferred are mixtures of TMS and TDS in a weight ratio of TMS to TDS of about 97:3 to about 20:80. These builders are disclosed in U.S. Patent 4,663,071, issued to Bush et al. on May 5, 1987.

Suitable ether polycarboxylates also include cyclic compounds, especially alicyclic compounds such as those described in U.S. Patents 3,923,679; 3,835,163; 4,158,635; 4,120,874 and 4,102,903.

Other useful detergent builders include the ether hydroxypolycarboxylates, which are represented by the structural formula:

HO-[C(R)(COOM)-C(R)(COOM-O]n-H

wherein M is hydrogen or a cation, the resulting salt being water soluble, preferably an alkali metal, ammonium or substituted ammonium cation; n is about 2 to about 15 (preferably n is about 2 to about 10, more preferably n has average values of about 2 to about 4); and each R is the same or different and is selected from hydrogen, C1-C4 alkyl or substituted C1-C4 alkyl (preferably R is hydrogen).

Other ether polycarboxylates include copolymers of maleic anhydride with ethylene or vinyl methyl ether, 1,3,5-trihydroxybenzene-2,4,6-trisulfonic acid, and carboxymethyloxysuccinic acid.

Organic polycarboxylate builders also include the various alkali metal, ammonium and substituted ammonium salts of polyacetic acids. Examples include the sodium, potassium, lithium, ammonium and substituted ammonium salts of ethylenediaminetetraacetic acid and nitrilotriacetic acid.

Also included are polycarboxylates such as mellitic acid, succinic acid, oxydisuccinic acid, polymaleic acid, benzene-1,3,5-tricarboxylic acid and carboxymethyloxysuccinic acid, and soluble salts thereof.

Citrate builders, e.g. citric acid and soluble salts thereof (especially the sodium salt), are polycarboxylate builders which are of great importance for liquid general purpose detergent formulations, but can also be used in granular compositions.

Other carboxylate builders include the carboxylated carbohydrates disclosed in U.S. Patent 3,723,322, Diehl, issued March 28, 1973.

Also suitable in the detergent compositions of the present invention are the 3,3-dicarboxy-4-oxa-1,6-hexanedioates and related compounds disclosed in U.S. Patent 4,566,984, Bush, issued January 28, 1986. Useful succinic acid builders include the C5 -C20 alkyl succinic acids and salts thereof. A particularly preferred compound of this type is dodecenyl succinic acid. Alkyl succinic acids typically have the general formula R-CH(COOH)CH2(COOH), i.e., are derivatives of succinic acid wherein R is a hydrocarbon, e.g. B. C₁₀-C₂₀ alkyl or alkenyl, preferably C₁₂-C₁₆, or wherein R may be substituted by hydroxyl, sulfo, sulfoxy or sulfone substituents, all as described in the above-mentioned patents.

The succinate builders are preferably used in the form of their water-soluble salts, including sodium, potassium, ammonium and alkanolammonium salts.

Specific examples of succinate builders include lauryl succinate, myristyl succinate, palmitylsuccinate, 2-dodecenyl succinate (preferred), 2-pentadecenyl succinate and the like. Lauryl succinates are the preferred builders of this group and are described in European Patent Application 200,263, published November 5, 1986.

Examples of useful builders also include sodium and potassium carboxymethyloxymalonate, carboxymethyloxysuccinate, cis-cyclohexane hexacarboxylate, cis-cyclopentane tetracarboxylate, water-soluble polyacrylates (these polyacrylates with molecular weights up to over about 2,000 can also be used effectively as dispersants), and the copolymers of maleic anhydride with vinyl methyl ether or ethylene.

Other suitable polycarboxylates are the polyacetal carboxylates disclosed in US Patent 4,144,226, Crutchfield et al., issued March 13, 1979. These polyacetal carboxylates can be prepared by contacting an ester of glyoxylic acid and a polymerization initiator under polymerization conditions. The resulting polyacetal carboxylate ester is then attached to chemically stable end groups to stabilize the polyacetal carboxylate against rapid depolymerization in alkaline solution, converted to the corresponding salt and added to a surfactant.

Polycarboxylate builders are also disclosed in U.S. Patent 3,308,067, Diehl, issued March 7, 1967. Such materials include the water-soluble salts of homo- and copolymers of aliphatic carboxylic acids such as maleic acid, itaconic acid, and methylenemalonic acid.

Other organic builders known in the art may also be used. For example, monocarboxylic acids and soluble salts thereof may be used with long chain hydrocarbyls. These would include materials commonly referred to as "soaps". Typically, chain lengths of C10-C20 are used. The hydrocarbyls may be saturated or unsaturated.

Other optional ingredients include soil release agents, chelating agents, clay soil removal/antiredeposition agents, polymeric dispersants, bleaching agents, brighteners, suds suppressors, solvents and aesthetic agents.

The detergent composition herein may be formulated in a variety of compositions, for example as laundry detergents as well as hard surface cleaners or dishwashing detergent compositions.

The compositions of the invention are further illustrated by the following examples.

Example I

The following compositions are prepared by combining the listed ingredients in the ratios indicated. In this example, one or more of the following peptide aldehydes are used:

Peptide aldehyde 1: CH3 O-(O)C-Phe-Gly-Ala-LeuH

Peptide aldehyde 2: CH3 N-(O)C-Phe-Gly-Ala-LeuH

Peptide aldehyde 3: CH3 O-(O)C-Phe-Gly-Ala-PheH

Peptide aldehyde 4: CH3 N-(O)C-Phe-Gly-Ala-PheH

Peptide aldehyde 5: CH3 SO2 -Phe-Gly-Ala-Leu-H

Peptide aldehyde 6: CH3 SO2 -Val-Ala-Leu-H

Peptide aldehyde 7: C6 H5 CH2 O(OH)(O)P-Val-Ala-Leu-H

Peptide aldehyde 8: CH3 CH2 SO2 -Phe-Gly-Ala-Leu-H

Peptide aldehyde 9: C6 H5 CH2 SO2 -Val-Ala-Leu-H

Peptide aldehyde 10: C6 H5 CH2 O(OH)(O)P-Leu-Ala-Leu-H

Peptide aldehyde 11: C6 H5 CH2 O(OH)(O)P-Phe-Ala-Leu-H

Peptide aldehyde 12: CH3 O(OH)(O)P-Leu-Gly-Ala-Leu-H.

Example II

The following formula is tested for % residual protease activity. Combinations of 0%, 0.1%, 0.2% and 0.3% Ca++ (from CaCl2) and 0%, 0.0006%, 0.00125% and 0.0025% peptide aldehyde (Synthesis Example 6) are used. The products are stored at 32.2ºC (90ºF) and assayed at weekly intervals for 42 days.

Component Wt. (%)

1,4-ethoxylated alkyl sulfate 30

Amine oxide 6

Polyhydroxy fatty acid amide 4

Non-ionic surfactant (C11E9) 5

Mg ion from MgCl₂ 1

Ca ion from CaCl2 see diagram below

Peptide aldehyde* see diagram below

Sodium xylenesulfonate 4

Solvent 6

Water to 100%

pH to 8

* Peptide aldehydes from synthesis example 6.

Example III

Results showing percent protease activity remaining after 42 days at 32.2ºC (90ºF). Using 0.01% Protease B enzyme.

Example IV

The following compositions are prepared by combining the listed ingredients in the proportions indicated.

1: x = degree of ethoxylation. The average degree of ethoxylation for the compositions is: A = 2.2; B = 0.6; C = 1.4; and D = 2.2.

2: Herein, the peptide aldehydes of Synthesis Example 6 are used.

Claims (1)

1. A liquid detergent composition comprising: (a) 1 to 95% by weight of the composition of a detersive surfactant; (b) an active proteolytic enzyme; (c) a source of calcium ions other than a compound of the formula RO(A)mSO3M, wherein R is an unsubstituted C10-C24 alkyl or hydroxyalkyl group having a C10-C24 alkyl moiety, A is an ethoxy or propoxy moiety, m is greater than zero and M is calcium; and (d) a peptide aldehyde of the formula: Z-B-NH-CH(R)-C(O)H wherein B is a peptide chain comprising 1 to 5 amino acid groups; Z is an N-capping group selected from the group consisting of phosphoramidate [(R"O)₂(O)-], sulfenamide [(SR")₂-], sulfonamide [(R"(O)₂S-], sulfonic acid [SO₃H], phosphinamide [(R")₂(O)P-], sulfamoyl derivative [R"O(O)₂S-], thiourea [(R")₂N(O)C-], thiocarbamate [R"O(S)C-], phosphonate [R"-P(O)OH], amidophosphate [R"O(OH)(O)P-], carbamate (R"O(O)C-) and urea (R"NH(O)C-), wherein each R" is independently selected from the group consisting of straight or branched, unsubstituted C₁-C₆ alkyl, phenyl, C₇-C₆ alkylaryl and cycloalkyl groups, wherein the cycloalkyl ring may span C₄-C₈ and may contain one or more heteroatoms selected from the group consisting of O, N and S, and R is selected from the group consisting of straight or branched, unsubstituted C₁-C₆ alkyl, phenyl and C₇-C₆ alkylaryl groups. 2. A liquid detergent composition according to claim 1, wherein the source of calcium ions is selected from calcium formate, calcium chloride, calcium acetate, calcium xylene sulfonate, calcium sulfate and mixtures thereof. 3. A liquid detergent composition comprising: a) 1 to 95% by weight of the composition of a detergent surfactant; b) an active proteolytic enzyme; c) a source of calcium ions selected from calcium formate, calcium chloride, calcium acetate, calcium xylene sulfonate, calcium sulfate and mixtures thereof; d) a peptide aldehyde having the formula: Z-B-NH-CH(R)-C(O)H wherein B is a peptide chain comprising 1 to 5 amino acid groups; Z is an N-capping group selected from the group consisting of phosphoramidate [(R"O)₂(O)-], sulfenamide [(SR")₂-], sulfonamide [(R"(O)₂S-], sulfonic acid [SO₃H], phosphinamide [(R")₂(O)P-], sulfamoyl derivative [R"O(O)₂S-], thiourea [(R")₂N(O)C-], thiocarbamate [R"O(S)C-], phosphonate [R"-P(O)OH], amidophosphate [R"O(OH)(O)P-], carbamate (R"O(O)C-) and urea (R"NH(O)C-), wherein each R" is independently selected from the group consisting of straight or branched, unsubstituted C1-C6 alkyl, phenyl, C7-C9 alkylaryl and cycloalkyl groups, wherein the cycloalkyl ring may span C4-C8 and may contain one or more heteroatoms selected from the group consisting of O, N and S; and R is selected from the group consisting of straight or branched, unsubstituted C1-C6 alkyl, phenyl and C7-C9 alkylaryl groups. 4. A liquid detergent composition according to claim 1 or 3, comprising a) 8 to 70% of the detergent surfactant; b) 0.0001% to 5% of an active proteolytic enzyme; c) 0.01% to 1% calcium ions; and d) 0.00001% to 5% of the peptide aldehyde. A liquid detergent composition according to claim 1 or 3, wherein the R groups are selected from the group consisting of methyl, iso-propyl, sec-butyl, iso-butyl, -C6H5-, -CH2-C6H5, and -CH2CH2-C6H5. 3. Liquid detergent composition according to claim 5, wherein the B peptide chains are selected from the group consisting of peptide chains having the amino acid sequences according to the general formula: Z-A⁵-A⁴-A³-A²-A¹-NH-CH(R)-C(O)H so that the following amino acids, if present, are: A¹ selected from Ala, Gly; A², if available, chosen from Val, Ala, Gly, Ile; A³, if present, selected from Phe, Leu, Val, Ile; A⁴, if present, any amino acid; A⁵, if present, any amino acid. 7. A liquid detergent composition according to claim 6, wherein the N-terminal end is protected by one of the N-capping group protecting groups selected from the group consisting of carbamates, ureas, sulfonamides, phosphonamides, thioureas, sulfenamides, sulfonic acids, phosphinamides, thiocarbamates, amidophosphates and phosphonamides. 8. A liquid detergent composition according to claim 7, wherein the N-terminal end of the protease inhibitor is protected by a methyl, ethyl or benzyl carbamate [CH3O-(O)C-; CH3-CH2O-(O)C-; or C6H5CH2O- (O)C-], methyl, ethyl or benzyl urea [CH3NH-(O)C-; CH3CH2NH- (O)C-; or C6H5CH2NH-(O)C-], methyl, ethyl or benzyl sulfonamide [CH3SO2-; CH3CH2SO2-; or C₆H₅CH₂SO₂-], and methyl, ethyl or benzylamidophosphate [CH₃O(OH)(O)P-; CH₃CH₂O(OH)(O)P-; or C₆H₅CH₂O(OH)(O)P-] groups. 9. A liquid detergent composition according to claim 8, wherein the petidaldehyde is selected from the group consisting of: CH₃SO₂Phe-Gly-Ala- Leu-H, CH₃SO₂Val-Ala-Leu-H, C₆H₅CH₂O(OH)(O)P-Val-Ala-Leu-H, CH₃CH₂SO₂-Phe-Gly-Ala-Leu-H, C₆H₅CH₂SO₂-Val-Ala-Leu-H, C₆H₅CH₂O(OH)(O)P-Leu-Ala-Leu-H, C6H5CH2O(OH)(O)P-Phe-Ala-Leu-H and CH3O(OH)(O)P-Leu-Gly-Ala-Leu-H. 10. A liquid detergent composition according to claim 9, wherein the source of calcium ions is partially replaced by a source of other divalent ions.