A DNA TRANSPORTER SYSTEM AND METHOD OF USE

Abstract: A DNA transporter system capable of non-covalently binding to DNA and facilitating the insertion of the DNA into a cell. The DNA transporter system includes a binding complex which non-covalently binds the DNA. The binding complex includes a molecule that is capable of non-covalently binding to the DNA and being covalently linked to a surface ligand and to a nuclear ligand. The surface ligand is capable of binding to a cell surface receptor and the nuclear ligand is capable of recognizing and transporting the transporter system through the nuclear membrane. A plurality of these binding complexes are attached to the DNA to facilitate the transport of the DNA into the cell. Additionally, a third binding complex which includes a virus can also be non-covalently linked to the DNA. The virus facilitates the movement of the DNA through the cytoplasm and into the nucleus. Also described are a variety of structures which can be used as part of the transporter system as well as methods of using the transporter system to introduce DNA into cells.

Description: A DNA TRANSPORTER SYSTEM AND METHOD OF USE The invention was partially supported by a grant from the United States government under HL-23741 awarded by the National Institute of Health. The government has certain rights in the invention. FIELD OF THE INVENTION The present invention relates generally to transporter systems for inserting DNA into the cell nucleus. It more particularly relates to targeting DNA into the nucleus of specific cells, both in vivo and in vitro. Additionally, it relates to methods of using lytic peptides to release DNA into the cellular interior. Further, it relates to the methods of using a transporter system to insert DNA into cells. BACKGROUND OF THE INVENTION Generally it has been recognized that it is desirable to introduce genes into specific types of cells as therapy for human disease. A targeted gene delivery system for nonviral forms of DNA and RNA requires 4 components: (a) a DNA or RNA molecule with a known primary sequence that contains the genetic information of interest, (b) a moiety that recognizes and bind to a cell surface receptor or antigen, (c) a DNA binding moiety and (d) a lytic moiety that enables the transport of the entire complex from the cell surface directly into the cytoplasm. Current gene delivery systems are asialoorsomucoid or transferrin, covalently linked to either a polymer of the amino acid lysine or ethidium homodimer and then complexed with DNA. The polylysine or the ethidium moiety provides a positively charged template for noncovalent binding of the negatively charged nucleic acid expression vector. After cellular uptake of these complexes, detectable, but not quantitatively significant, amounts of the reporter gene have been found. The principal limitation of this approach is the inability to prepare reproducible proteinaceous complexes in a consistent manner for gene delivery. The actual site of ligation is unknown. The covalent coupling of polylysine or the dye with proteins is nonspecific and gives a random mixture of conjugates. Because the conformation of positively charged surfaces is formed by chance, binding of DNA to the charged template is also variable. The high molecular weight and variable stoichiometry of the components of the complex has made it difficult to prepare the complexes either consistently well or in sufficient quantity for in vivo delivery. The mixture of compounds precludes a molecular definition of the biologically active reagent and present formulations are inadequate for this reason. The present invention is an improved delivery system using chemically defined template molecules. It provides an effective and efficient method to target gene delivery to specific cells. It further provides an effective and efficient method to release DNA into the cellular interior after the DNA has been internalized by the use of lytic peptides covalently linked to template molecules through an acid sensitive linker. It can also be used to target cells to the nucleus of the specific cells. SUMMARY OF THE INVENTION An object of the present invention is a transporter system capable of efficiently inserting DNA into the nucleus of a cell. An additional object of the present invention is a compound useful as a DNA transporter system. A further object of the present invention is a method of inserting genes into cells using a DNA transporter system. Another object of the present invention is a method of releasing genes into the cellular interior after the DNA has been internalized by the use of lytic peptides covalently linked to template molecules through an acid sensitive linker. Thus in accomplishing the foregoing objects, there is provided in accordance with one aspect of the present invention a DNA transporter system for inserting specific DNA into a cell comprising a plurality of a first DNA binding complex, said complex including a first binding molecule capable of non-covalently binding to DNA, said first binding molecule covalently linked to a surface ligand, said ligand capable of binding to a cell surface receptor; a plurality of a second DNA binding complex, said complex including a second binding molecule capable of non-covalently binding to DNA, said second binding molecule covalently linked to a nuclear ligand, said nuclear ligand capable of recognizing and transporting the transporter system through a nuclear membrane; wherein said plurality of first and second DNA binding complexes are capable of simultaneously, non-covalently binding to the specific DNA. Specific embodiments the DNA transporter system of the present invention can further include a plurality of a third DNA binding complex, said complex including a third binding molecule capable of non-covalently binding to DNA, said third binding molecule covalently linked to a virus or a lytic peptide; wherein said plurality of third DNA binding complexes are capable of simultaneously, non-covalently binding to the specific DNA. In additional embodiments of the present invention, the binding molecules can be selected from the group consisting of spermine, spermine derivative, histones and polylysine. In the preferred embodiment, a spermine derivative is used. In specific embodiments of the present invention, the surface ligand is a molecule which binds to a receptor selected from the group consisting of a folate receptor, biotin receptor, lipoic acid receptor, low density lipoprotein receptor, asialoglycoprotein receptor, Fab' fragment of IgG, insulin-like growth factor type II/cation-independent mannose-6-phosphate receptor, calcitonin gene-related peptide receptor, insulin-like growth factor I receptor, nicotinic acetylcholine receptor, hepatocyte growth factor receptor, endothelin receptors or bile acid receptor. These receptors function in most, if not all, cell types, although their abundance varies considerably with cell type. Organ specificity can be achieved either by direct injection into the organ of interest or by intra-arterial administration with a venous outflow to remove any material not taken up by the tissue. In the preferred embodiment, the asialoglycoprotein receptor is used. In the preferred embodiment, a spacer molecule is also used to link the binding complex to the surface ligand, the binding complex to the nuclear ligand and the binding complex to the virus. The spacer molecule is usually hydrophilic and ranges from 6 to 30 carbons. In the preferred embodiment, a compound from 6 to 16 carbons is used. A further embodiment is a spacer molecule that is polymer of [ (gly) j (ser) j ] k where i ranges from 1 to 6, j ranges from 1 to 6, and k ranges from 3 to 20. The first, second and third DNA binding complexes can be the same common DNA binding complex where any combination of cell surface ligand, nuclear ligand, virus or lytic peptide can be linked to the same binding molecule. Additional embodiments of the present invention include specific compounds which can be used as ligands in the transporter system. Further embodiments of the present invention include methods of using the transporter system introducing DNA into cells; in vivo and in vitro targeting of the insertion of DNA into specific cells; prevention and treatment of disease; and modification of animals. Other and further objects, features and advantages will be apparent from the following description of the presently preferred embodiments of the invention, which are given for the purposes of disclosure when taken in conjunction with the company drawings. BRIEF DESCRIPTION OF THE DRAWING Figures 1-3 show the schematic synthesis of the receptor ligands. Figure 4 shows a schematic synthesis of the virus ligand. Figures 5-13 show a schematic flow chart for the synthesis of the DNA transporter system. Figure 14 is a schematic diagram of insertion of a triplex forming oligonucieotide or peptidyl nucleic acid by attachment of a ligand. Figure 15A is a schematic representation of using a ligand to target to a triplex forming oligonucieotide to a duplex DNA. Figure 15A is a schematic representation using a ligand to target a triplex forming peptidyl oligonucieotide to a duplex DNA. Figure 15B is a specific ligand for targeting muscle. Figure 15C shows analogs useful in the compound of 15A. Figure 15D is an analog of 15A. Figure 16 shows a specific targeting of a ligand for the SV40 sequences. Figure 17 shows the targeting with a ligand of a sequence to the c-myc promoter region. Figure 18 is a schematic of the peptide ligands and other ligands and shows the abbreviations used herein for the ligands. Figure 19 shows the schematic synthesis of receptor ligands. Figure 20 shows specific ligands for targeting to muscle. Figure 21 shows the folyl-spermine derivative. Figure 22 shows a specific ligand for targeting muscle. Figure 23 is a schematic of the peptide ligands and other ligands and shows the abbreviations used herein for the ligands. Figure 24 is a schematic representation of a synthetic route for bifunctional acid sensitive linkers. Figure 25 is a schematic representation of a trimeric fusogenic peptide. Figure 26 is a schematic representation of the synthetic route for a trimeric fusogenic peptide. Figure 27 is a schematic representation of a synthetic route for a monomeric fusogenic peptide. Figure 28 is a schematic representation of a synthetic route for a trimeric fusogenic peptide. Figure 29 is a schematic representation of a synthetic route for a trimeric fusogenic peptide. The drawings are not necessarily to scale. Certain features of the invention may be exaggerated in scale or shown in schematic form in the interest of clarity and conciseness. DETAILED DESCRIPTION OF THE INVENTION It will be readily apparent to one skilled in the art that various substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention. The term "DNA transporter" refers to a molecular complex which is capable of non-covalently binding to DNA and efficiently transporting the DNA through the cell membrane. Although not necessary, it is preferable that the transporter also transport the DNA through the nuclear membrane. The term "spacer" as used herein refers to a chemical structure which links two molecules to each other. The spacer normally binds each molecule on a different part of the spacer molecule. In the present invention, the spacer is a hydrophilic molecule comprised of about 6 to 30 carbon atoms. In the preferred embodiment, it usually contains 6 to 16 carbons. In a second embodiment, the spacer is a hydrophilic peptide, a polymer of [ (gly) j (ser) j ] k where i ranges from 1 to 6, j ranges from 1 to 6, and k ranges from 3 to 20. A receptor is a molecule to which a ligand binds specifically and with relatively high affinity. It is usually a protein or glycoprotein, but may also be a glycolipid, a lipopolysaccharide, a glycosaminoglycan or a glycocalyx. For this patent, the epitope to which an antibody or its fragment binds is construed as a receptor, since the antigen:antibody complex undergoes endocytosis. The term "cell surface receptor" as used herein refers to a specific chemical grouping on the surface of a cell for which a ligand can attach. The receptor facilitates the internalization of the ligand and attached molecules. Cell surface receptors which have been found to be useful in the present invention include the folate receptor, the biotin receptor, the lipoic acid receptor, the low density lipoprotein receptor, the asialoglycoprotein receptor, IgG antigenic sites, insulin-like growth factor type II/cation-independent mannose-6- phosphate receptor, calcitonin gene-related peptide receptor, insulin¬ like growth factor I receptor, nicotinic acetylcholine receptor, hepatocyte growth factor receptor, endothelin receptor, bile acid receptor. The term "lytic peptide" refers to a chemical grouping which penetrates a membrane such that the structural organization and integrity of the membrane is lost. As the result of the presence of the lytic pepitde, the membrane undergoes lysis, fusion or both. Lytic peptides which have found to be useful in the present invention include those of the Othromyxoviridae, Alphaviridae, and Arenaviridae. The term "nuclear receptor" refers to a chemical grouping on the nuclear membrane which will bind a specific ligand and help transport the ligand through the nuclear membrane. Nuclear receptors which have been found to be useful in the present invention include the receptors which bind the nuclear localization sequences. As used herein, the term "ligand" refers to a chemical compound or structure which will bind to a receptor. In the present invention, useful cell surface ligands include folate (A Figure 1), lipoic acid (G Figure 3), biotin (B Figure 2), apolipoprotein E sequence (Pep2 Figure 18), D (Figure 18), Asp (bis-LacAHT) E (Figure 18), Fab' (Figure 18), Compound P L-tyrosyl-L-aspartoyl-Jbis-{N-[6-[[6-O-phosphoryl-α-D- mannopyranosyl]oxy]hexyl]-L-alaninamide] (Figure 19), Compound J 3-{N- [3,4,5-tris-(2-triethylammoniumethoxy)benzoic acid, For Peptides Pepl2- 19, X is the 3(2-pyridyldithio)propionyl moiety (Figure 20), Pepl2. [Gin 0 , Leu 27 , e-X-Lys 67 ]-insulin-like growth factor II, Pepl3. [Gin 0 , e-X- Lys 3 , Leu 27 , Arg 67 ]-insulin-like growth factor II, Pepl4. Y°-e-X-Lys 24 - calcitonin gene-related peptide, Pepl5. [Asu 2,7 , Y 8 , e-X-K 24 ]-calcitonin gene-related peptide, Pepl6. (Gin 0 , Leu 27 , e-X-Lys 54 , Arg 55 , Arg 67 )- insulin-like growth factor II, Pepl7. ta-X-des- ( l-3 ) - {Arg 65 , Arg 67 ]- insulin-like growth factor I, Peplδ. e-X-K°-thymopoietin, Pepl9. e-X-K 4 - thymopoietin, 7α,12α-dihydroxy-3β-(ω-aminoalkox )-5-β-cholan-24-oic acid, Pep20 hepatocyte growth factor, Pep21 endothelin-1, Pep22 N- succinyl-[glu 9 r ala u,15 ]-endothelin-l(8-21), Pep23 r-atrial natriuretic factor (99-126). In the preferred embodiment, E is used for liver and P is used for the muscle. One skilled in the art will readily recognize that the ligand chosen will depend on which receptor is being bound. Since different types of cells have different receptors, this provides a method of targeting DNA to specific types of cells, depending on which cell surface ligand is used. Thus, the preferred cell surface ligand may depend on the targeted cell type. In the present invention, useful lytic peptides are Pep24 influenza GLFEAIAGFIEDGWEGMIDGGGC, Pep25 SFV El KVYTGVYPFMWGGAYCFCD, and Pep26 Lassa gp2 GGYCLTRWMLIEAELKCFGNTAV. In the present invention, useful nuclear ligands are shown on Figure 18 and include Pep3, Pep4, Pep5, Pep6, Pep7, Pep8, Pep9 and PeplO. One embodiment of the present invention is a DNA transporter system for inserting specific DNA into a cell comprising a plurality of a first DNA binding complex, said complex including a first binding molecule capable of non-covalently binding to DNA, said first binding molecule covalently linked to a surface ligand, said surface ligand capable of binding to a cell surface receptor; a plurality of a second DNA binding complex, said complex including a second binding molecule capable of non-covalently binding to DNA, said second binding molecule covalently linked to a nuclear ligand, said nuclear ligand capable of recognizing and transporting a transporter system through a nuclear membrane; wherein said plurality of first and second DNA binding complexes are capable of simultaneously, non-covalently binding to a specific DNA. Alternative embodiments of the present invention further included a plurality of a third DNA binding complex, said complex includes a third binding molecule capable of non-covalently binding to DNA, said third binding molecule covalently linked to a virus or lytic peptide; wherein said plurality of third DNA binding complexes are capable of simultaneously, non-covalently binding to a specific DNA. The first binding molecule, the second binding molecule and third binding molecule can each be selected from the group consisting of spermine, spermine derivative, histones, cationic peptides and polylysine. Spermine derivative refers to analogues and derivatives of spermine and include compounds IV, VII, XXI, XXXIII, XXXVI, LIV, LVI, LXXXII, LXXXIV and CX. In the DNA transporter system, the first, second and third binding molecules can be different or can be the same. In the preferred embodiment, the first, second and third binding molecules are the same and are preferably a spermine derivative, most preferably D. When the transporter enters the cell it is engulfed by an endosome. A virus or lytic peptide can be used to break down the endosomal membrane, freeing the transporter into the cytoplasm. The virus is usually selected from the group consisting of adenovirus, parainfluenza virus, herpes virus, retrovirus and hepatitis virus. In one preferred embodiment, the adenovirus of the structure F Figure 4 is used. The lytic peptide is selected from the peptides from proteins of the Othromyxoviridae, Alphaviridae, and Arenaviridae. In one preferred embodiment, the peptide of the structure Pep24. The surface ligand, the nuclear ligand and the virus or lytic peptide can be attached directly to the first, second, and third binding molecules respectively; however, in the preferred embodiment, there is a spacer between the binding molecules and the ligands or virus. Some examples of binding molecules containing spacers which are useful in the present invention include compounds XI, XII, XL, XLI, LX, LXI, LXXXVIII, XC, CXVIV, CXVI, XV, XVI, XVIII, XXI, XLV, LXVII, ΣLVII, L, LXV, LXX, XCIV, XCVI, XCIX, XXIV, XXV, LXXIII, LXXIV, CII, CV. Another embodiment of the DNA transport system for inserting specific DNA into a cell comprises a plurality of a common DNA binding complex. Each of said complexes includes a binding molecule capable of non-covalently binding to DNA, said binding molecule is attached to both a surface ligand capable of binding to a cell surface receptor and a nuclear receptor capable of recognizing and transporting the transporter system through a nuclear membrane. In preferred embodiments, this DNA transporter also includes at least one spacer which links the surface ligand and nuclear ligand to the binding molecule. Further embodiments of this DNA transporter include a virus, lytic peptide or Fab' also attached to the common binding molecule. Again, either a virus or lytic peptide can be attached by the same or a different spacer. The DNA transporter of the present invention can be used in a variety of methods for introducing DNA into the cell. One method comprises contacting the DNA with the elements of the transporter such that the transporter elements non-covalently bind to the DNA. This DNA transporter system is contacted with the cell for insertion. Another method involves the in vitro targeting of the insertion of DNA into a cell, In this procedure the DNA transporter system is contacted with the cell, however, the transporter has a cell surface ligand which is cell type specific. The invention can also be used for prevention and treatment of disease by introducing a therapeutic dose of DNA into the cells by contacting the DNA transporter system with the cells to be treated. This treatment, therapy and prevention can be used in both humans and in animals. Another embodiment of the present invention is a method of modifying the genetic makeup of animals by introducing DNA into the cells of the animals. This can be used to modify production in domestic animals. To make the DNA binding asialoglycoprotein receptor (ASGPR) Ligand, a derivative of the naturally occurring DNA binding ligand spermine is synthesized. This spermine derivative can be covalently linked to a variety of cell specific ligands. The chemical structure of the spermine analog is IV. A spermine analog is coupled to a high affinity ligand for the ASGPR. The highest affinity for the ASGPR is achieved with two clusters of three galactose groups separated by an aspartic acid backbone. This structure Asp(bis-LacAHT) is E. The spermine derivative and the Asp(bis-LacAHT) are the two component molecules which are coupled to give the transporter. This compound is used to define the structural features necessary for high efficiency in vivo delivery of the expression vector. One skilled in the art will recognize that this is only one of many compounds from which second or third generation products are derived. This DNA binding spermine derivative has a wide variety of applicability in gene delivery systems for a variety of cell types of interest (for example liver, muscle, endothelium and skin) as long as the specific cell surface receptors are present for targeting. For cell specific target of expression vectors in vivo the system includes three components which are comprised of a DNA expression vector, a spermine derivative which binds the DNA to achieve a cell specific targeting, and pH sensitive liposomes or lytic peptides with a cell specific ligand to release the vector into the cytoplasm of the cell. The combination of these three components are taken up by the cell containing the receptors for both of the high affinity ligands. After co-internalization of the vector ligand complex and the lytic agent throughout the same coated pit on the plasma membrane of the cell, the decrease in pH that occurs immediately after endosome formation causes spontaneous lysis of the endosome. The vector is released into the cytoplasm for transfer to the nucleus where the targeted gene is expressed. The pH sensitive liposomes have a second ligand specific for the same cell type as the DNA binding ligand. With the hepatocytes a ligand containing two copies of the apolipoprotein "E" sequence ("APO") Pep2 is used. In vivo, this ligand has high affinity for the low density lipoprotein receptor related protein on the.hepatocyte plasma membrane. The following examples are offered by way of illustration and are not intended to limit the invention in any manner. Example 1 Synthesis of Components of Receptor ligands Examples of the specific components of the receptor ligands are shown in Figure 18 and Figures 1-4. A. In Figures 1-3 the schematic synthesis of A, B, G, P, J and M are shown. The actual synthesis for A, B, G, P, J and M are very similar. As an example, for the preparation of A, dissolve 1 mmol of folic acid in 2 mL dry dimethylformamide (DMF), add 1.3 mmol l-ethyl-3- [3-(dimethylamino)propyl)carbodiimide and stir in the dark under N 2 overnight at 4°, then add 1.3 mmol N-hydroxysuccinimide with stirring continued for another 6 hr at 4°. Add bis(2-aminoethane)disulfide, 1 mmol in 0.5 mL dry dimethylformamide, dropwise to the reaction mixture and stir for an additional 4 hr. Add 15 mL water to precipitate the product. After centrifugation, the precipitate is washed and dissolved in oxygen-free 0.1 M NH 4 0H. This solution is applied to an anion exchange resin and equilibrated in degassed 0.05 M NH 4 C0 3 containing 20% acetonitrile. The γ-isomer is separated from unreacted starting materials and the -isomer by chromatography in 0.1 M NH 4 CO s containing 20% acetonitrile. The appropriate fractions are pooled and lyophilized to obtain the product. The synthesis of the nicotinic acetylcholine receptor ligand, component J, 3-{N-[3,4,5-tris-(2-triethylammoniumethoxy)benzoic acid is the same as for folic acid, component A except the 1 mmol of folic acid 2 mL dry DMF is replaced with 1 mmol 3-{N-[3,4,5- tris-(2- triethylammoniumethox )benzoic acid in 2 mL dry DMF. The synthesis of the nicotinic acetylcholine receptor ligand, component J, 3-{N-[3,4,5- tris-(2-triethylammoniumethoxy)benzoic acid is the same as for folic acid, component A except the 1 mmol of folic acid 2 mL dry DMF is replacedwith 1 mmol 3-{N-[3,4,5-tris-(2-triethylammoniumethoxy)benzoic acid in 2 mL dry DMF. This compound can be further reacted to yield A' . Dissolve 2 mmol A in 10 mL oxygen-free 0.01 M NH 4 CO s containing 2 mmol dithioerythritol, stir for 2 hr. The solution is applied to an anion exchange resin equilibrated in degassed 0.1 M NH 4 C0 3 containing 20% acetonitrile. The reduced folate derivative is separated from unreacted starting materials by chromatography in 0.1 M NH 4 C0 3 containing 20% acetonitrile. The appropriate fractions are pooled, lyophilized, and then dissolved in 10 mL dry dimethylformamide for dropwise addition to a vigorously stirred solution of 2,2'-dipyridinedisulfide, 4 mmol dissolved in 10 mL ethanol containing 0.4 mL glacial acetic acid. After overnight at room temperature protected from light, the solvent is removed in vacuo. Add degassed 0.1 M NH 4 C0 3 to effect solution and then chromatograph as before to obtain the desired product. Both the original A, B and G and the further reacted A' , B' and G' have been used. The synthesis of the nicotinic acetylcholine receptor ligand, component J, 3-{N-[3,4,5-tris-(2-triethylammoniumethoxy)benzoic acid is the same as for folic acid, component A except the 1 mmol of folic acid 2 mL dry DMF is replaced with 1 mmol 3-{N-[3,4,5-tris-(2- triethylammoniumethoxy)benzoic acid in 2 mL.dry DMF. One skilled in the art will recognize that other vitamins and analogs of these vitamins can be used. Since the different vitamins and analogs will have different affinities, uptake and selectivity for the membrane receptors, the specific vitamin or analog is chosen to maximize the specificity and uptake. B. The peptides including Pepl through Pepll and Pep21 through 26 can be synthesized by a variety of methods. In the present invention solid phase synthesis on a support is preferred, except for peptides 12, 13, 16, 17 and 20 which are recombinant proteins produced by expression vectors in bacteria, yeast, baclovirus or mammalian systems. Peptides Pepl through Pep6 and Pepl2 through 23 are examples of peptides or peptide analogs. These peptides target and bind to membrane receptors. One skilled in the art recognizes that other peptides or analogs to other membrane receptors can be used, and that the order of the amino acid sequence can be reversed, inverted and/or repeated, while still maintaining the transporter characteristics. The selection of a specific peptide will depend on the tissue and membrane receptor which is targeted. By selecting specific peptides, one skilled in the art recognizes the binding efficiency, uptake and specificity can be regulated. This can be used for tissue specificity. Peptides Pep24 through Pep26 are examples of peptides or peptide analogs. These peptides lyse membranes. One skilled in the art recognizes that other peptides or analogs to other lytic peptides can be used, and that the order of the amino acid sequence can be reversed, inverted and/or repeated, and still maintain the lytic characteristics. The selection of a specific peptide will depend on the tissue and membrane receptor which is be targeted. I. SPERMINE TEMPLATE a. nonomeric fusogenic peptide covalently linked to a polycation through an acid sensitive, reducible spacer. NH 2 - ( Ctt, ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 "NH- ( CH 2 ) 3 -NH 2 OC^CONH-CH g C^-NH-R ( 9 ) where R = Pep24-SS-CH 2 CH 2 NH-α-CO-aconityl-γ-CO- H g N- ( CH 2 ) 3 -NH-CH 2 -CH- ( 0 ) 2 ~NH- ( C&, ) 3 -NH 2 ( IV) OH + C 15 H u OCO-Cl I FMOC FMOC FMOC-NH- ( CH 2 ) g -N I-CH- 2 -CH- ( OL, ) 2 "N I- ( Ci^ ) 3 -NH-FMOC ( 1 ) OH + BrCH 2 COOCH 3 , NaOCH 3 1 FMOC FMOC FMOC-NH-(CH 2 ) 3 - I-CH 2 -CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NH-FMOC (2) OCH 2 COOCH 3 + OH ~ 4. + HJNCHJCHJJN^ ( 4 ) l + 6 FMOC FMOC FMOC-NH- ( CR, ) 3 -N I-C& 2 -CH- (CR,) 2 -N I- (CH 2 ) 3 -NH-FMOC (7 ) OCH 2 CONHCH 2 CH 2 NH-γ-CO-cis-C 4 H 4 0 2 -CONH ( CH j .) 2 SS-C S H 4 N 4- + piperidine H 2 N-(CH 2 ) 3 -NH-CH 2 -CH-(CH 2 ) 2 -NH-(CH 2 ) 3 - H 2 (8) OCI^CONH^CH^-γ-CO-σis-C^O g -CONH ( CR-j 2 SS-C 5 H 4 N 1 + influenza fusion peptide GLFEAIADFIENGWEGMIDGGGC-SH (Pep24) NH 2 -(CH 2 ) 3 -NH-CH 2 -CH-(CH 2 ) 2 -NH-(CH 2 ) 3 -NH 2 OCH 2 CONH-CH 2 CH 2 -NH-R (9) Where R = GLFEAIADFIENGWEGMIDGGGC-SS-CH 2 CH 2 NH-α-CO-aconityl-γ-CO- Detailed Preparative Procedures. Combine 2 mmol (S)-hydroxyspermine (IV), 4 mmol 4- pyrrolidinopyridine and 4.1 mmol 9-fluorenyImethy1 chloroformate in 40 mL anhydrous benzene and stir overnight at room temperature under N 2 . Separate the desired product (1) by solid phase extraction on phenyl- silica and elution with a linear gradient of ethyl acetate to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product as an amorphous solid. Add 4 mL of dry benzene to 1 mmol Jr^CO g and 2 mmol 18-crown-6 and stir for 20 min. Add 2 mmol 1 in 4 mL of dry benzene, followed by 2 mmol methyl bromoace ate in 2 mL benzene. After 4 hr, add 25 mL of water and extract with 3 portions of 25 mL benzene. Remove the solvent in vacuo and dissolve the residue in 10 mL ethanol containing 2 mmol potassium hydroxide. After overnight at room temperature, the solution is transferred to a separatory funnel, to which 2 mmol of HCl, 5 mL of water and 25 mL of benzene is added. After extraction with 3 additional portions of benzene, the combined organic phase is tc en to dryness in vacuo, redissolved in a minimum volume of ethyl a state, diluted with enough petroleum ether to create slight turbidity and cooled at 4° to promote crystallization of 3. Dissolve 1 mmol of 3 in 2 mL dry dimethylformamide, add 3.0 mmol l-ethyl-3-[3-(dimethylamino)propyl)carbodiimide and stir 2 hr, then add 1.1 mmol N-hydroxysuccinimide and continue stirring for another 6 hr at room temperature. This solution is added dropwise to 10 mmol of 1,2- diaminoethane in 0.5 mL dry dimethylformamide, and stirring continued for an additional 4 hr, when the reaction is complete as monitored by thin layer chromatography. The solution is applied to an cation exchange resin equilibrated in degassed water. The product is separated from unreacted starting materials by a gradient from 0.0005 to 2.0 M HCl. The appropriate fractions are pooled and lyophilized to obtain the product (4). The Synthetic route for the bifunctional acid sensitive linker (6) is shown in Figure 24. Combine 10 mmol cis-aconityl anhydride 10 mmol with 2-(2'- aminoethyldithio)-pyridine in 20 mL dry DMF under N 2 protected from light at room temperature for 18 hr. Remove the solvent in vacuo, dissolve the residue in the minimum amout of 3M NH 4 C0 3 , dilute 10 fold and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 C0 3 and elute with a gradient to 1.0M NH 4 C0 3 . The appropriate fractions are pooled, lyophilized and the residue (5) dissolved in 2- propanol and crystallized at 4° after the solution is made turbid by the addition of diethyl ether. The product (5) is collected by filtration. Dissolve 1 mmol of 5 in 2 mL dry DMF, add 1.1 mmol N- hydroxysuccinimide and 1.1 mmol dicyclohexylcarbodiimide and continue stirring for another 24 hr at 4°. Remove the solvent in vacuo, dissolve the residue in the minimum amout of 2-propanol and crystallize at 4° after the solution is made turbid by the addition of diethyl ether. The product (6) is collected by filtration. Dissolve 1 mmol of 6 in 2 mL dry DMF, add 1.1 mmol N- hydroxysuccinimide and 1.1 mmol dicyclohexylcarbodiimide and continue stirring for another 6 hr at 4°. One mmol of 4 in 0.5 mL dry dimethylformamide, is added dropwise to the preceding reaction mixture and stirring continued for an additional 4 hr. Remove the solvent in vacuo and dissolve the residue in 10 mL 50% piperidine in DMF (v/v) . Again remove the solvents in vacuo, solubilize the residue (7 ) in 0.1 M NH 4 OH, and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 CO a containing 20% acetonitrile and eluted with a gradient of 0 to 0.25 M acetonitrile in 0.1 M NH 4 CO s . The appropriate fractions are pooled and lyophilized to obtain the product (8) . To a stirred solution of 0.5 mmol of lytic peptide (Pep24) in 5 mL PBS, pH 7.4, at 4°, add 0.1 mmol of 8 in PBS dropwise. After 18 hr, chromatograph over a molecular sieve to separate the product (9) from unreacted starting materials. b. trimeric fusogenic peptide covalently linked to a polycation through an acid sensitive, reducible spacer. NR,-(CR.J g -NH-CR.-CH-(CR,) 2 -NH-(CR,) 3 -NR. OCR,CONH-CH 2 CH 2 -NH-R ( 16 ) where R = -α-CO-aconityl-γ-COHNC- (OCH 2 CH 2 NHCO ( CH 2 ) 5 NH-R ' ) 3 and R' = Pep24-SS-CH 2 CR,CO- The Series II synthetic route (New compounds 10 - 16) is shown in Figure 25. Detailed procedures. Add 4 mL of dry benzene to 1 mmol 1^00 3 and 2 mmol 18-crown-6 and stir for 20 min. Add 2 mmol N-t-BOC- tris-(hydroxymethyl)methane in 4 mL of dry benzene, followed by 20 mmol l-bromo-2-(N-5-FMOC- aminohexanoyl)-aminoethane in 2 mL benzene. After 4 hr, add 25 mL of water and extract with 3 portions of 25 mL benzene. Remove the solvent in vacuo and dissolve the residue (10) in 10 mL 50% piperidine in DMF (v/v) . After 6 hr, remove the solvents in vacuo, solubilize the residue in 20 mL ethyl acetate, wash with water until neutral and dry over molecular sieve before solid phase extraction and chromatography on octadecyl-silica, using a gradient of acetonitrile to 100%. Pool the appropriate fractions to obtain 11. Combine 2 mmol 11 in 20 mL acetonitrile with 7 mmol of succinimidyl 3(2-pyridyldithio)-propionate in ethanol. After 60 min, dilute with sufficient water to create a slight turbidity and apply to octadecyl-silica, again using a gradient of acetonitrile from 0 to 100%. Pool the appropriate fractions to obtain 12. Dissolve 1 mmol 12 in 20 mL 3N HCl at 4° and allow to stand for 60 min before the solution is taken to dryness in vacuo. The residue is resuspended in water and solublized with the minimum amount of acetonitrile and chromatographed on octadecyl-silica, again using a gradient of acetonitrile from 0 to 100%. Pool the appropriate fractions to obtain 13. Dissolve 1 mmol of 13 in 2 mL dry DMF, add 3 mmol cis-aconityl anhydride and stir under N 2 overnight at 4°. Remove the solvents in vacuo, solubilize the residue in 0.1 M NH 4 OH, and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 C0 3 containing 20% acetonitrile and eluted with a gradient of 0 to 0.25 M acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product (14). Dissolve 1 mmol of 14 in 2 mL dry DMF, add 1.1 mmol N- hydroxysuccinimide and 1.1 mmol dicyclohexylcarbodiimide and continue stirring for another 6 hr at 4°. One mmol of 4 in 0.5 mL dry dimethylformamide, is added dropwise to the preceding reaction mixture and stirring continued for an additional 4 hr. Remove the solvent in vacuo and dissolve the residue in 10 mL 50% piperidine in DMF (v/v). Again remove the solvents in vacuo, solubilize the residue in 0.1 M NH 4 OH, and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 C0 3 containing 20% acetonitrile and eluted with a gradient of 0 to 0.25 M acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product (15). To a stirred solution of 0.5 mmol of lytic peptide (Pep24) in 5 mL PBS, pH 7.4, at 4°, add 0.1 mmol of 15 in PBS dropwise. After 18 hr, chromatograph over a molecular sieve to separate the product (16) from unreacted starting materials. c. trimeric fusogenic peptide covalently linked to a polycation through an acid sensitive, reducible spacer. NR j -(CR j ) 3 -NH-CR j -CH-(CR,) 2 -NH-(CR,) 3 -N , OCH 2 CONH-CH 2 CH 2 -NH-R" where R" --------- ε-R' e-R' e-R' -β-C0-aconϊtyl-γ-C0HN-glu-ly Is-gly-lyIs-gly-pro-gly-gly-lyIs-β,γ-diamfnobutyπ ' c-C0NH 2 (20) with the γ-carboxyl of glu 1 in amide linkage with the γ-amino of α,γ- diaminobutyric acid 10 and R' = Pep24-SS-CH 2 CH 2 CO- The Series III synthetic route (New compounds 17 - 20) is shown in Figure 26. Detailed experimental procedures. Solid phase peptide synthesis with conventional reagents and procedures gives 17. If is obvious to one skilled in the art that homologs of 2,4-diaminobutyric acid, such as orinithine and lysine, could be substituted for this residue and that other amino acids, such as serine, alanine, and aspartic acid, could be substituted for gly. Dissolve 1 mmol of 17 in 2 mL dry DMF, add 3 mmol σis-aconityl anhydride and stir under N 2 overnight at 4°. Remove the solvents in vacuo, solubilize the residue in 0.1 M NH 4 0H, and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 CO s containing 20% acetonitrile and eluted with a gradient of 0 to 0.25 M acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product (18) . Dissolve 1 mmol of 18 in 2 mL dry DMF, add 1.1 mmol N- hydroxysuccinimide and 1.1 mmol dicyclohexylcarbodiimide and continue stirring for another 6 hr at 4°. One mmol of 4 in 0.5 mL dry dimethylformamide, is added dropwise to the preceding reaction mixture and stirring continued for an additional 4 hr. Remove the solvent in vacuo and dissolve the residue in 10 mL 50% piperidine in DMF (v/v) . Again remove the solvents in vacuo, solubilize the residue in 0.1 M NH 4 OH, and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 C0 3 containing 20% acetonitrile and eluted with a gradient of 0 to 0.25 M acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product (19). To a stirred solution of 0.5 mmol of lytic peptide (Pep24) in 5 mL PBS, pH 7.4, at 4°, add 0.1 mmol of 19 in PBS dropwise. After 18 hr, chromatograph over a molecular sieve to separate the product (20) from unreacted starting materials. a. monomeric fusogenic peptide covalently linked to a polycation through an acid sensitive, reducible spacer. tyr-lys-ala-(lys^trp-lys-CON^ NHCO-CR J -CR J -CONH-CR J CR J -NH-R (24) where R = Pep24-SS-CH 2 CH 2 NH-α-CO-aconityl-γ-CO- The Series IV synthetic route (New compounds 21 - 24) is shown in Figure 27. Detailed experimental procedures. Solid phase peptide synthesis with conventional reagents and procedures gives 21. Dissolve 1 mmol of 21 in 2 mL dry DMF, add 3 6 and stir under N 2 overnight at 4°. Remove the solvents in vacuo, solubilize the residue in acetonitrile and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 CO s containing 20% acetonitrile and eluted with a gradient of 0 to 1.0 M acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product (22) . Dissolve 1 mmol of 22 in 10 mL 50% piperidine in DMF (v/v) . Remove the solvents in vacuo, solubilize the residue in 0.1 M NH 4 OH, and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 CO s and elute with a gradient to 1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product (23). To a stirred solution of 0.5 mmol of lytic peptide (Pep24) in 5 mL PBS, pH 7.4, at 4°, add 0.1 mmol of 23 in PBS dropwise. After 18 hr, chromatograph over a molecular sizing column to separate the product (24) from unreacted starting materials. b. trimeric fusogenic peptide covalently linked to a polycation through an acid sensitive, reducible spacer. tyr-lys-ala-(lys)^trp-lys-CONR. NHCO-CH 2 -CH 2 -CO--rø-CH 2 CH 2 -NH-R (27) where R = -α-CO-aconityl-γ-COHNC-(OCH 2 CJ^NHCO(Cl^) 5 NH-R' ) 3 and R' = Pep24-SS-CH 2 CH 2 CO- The Series V synthetic route (New compounds 25 -27) is shown in Figure 28. Detailed experimental procedures. Dissolve 1 mmol of 14 in 2 mL dry DMF, add 1.1 mmol N- hydroxysuccinimide and 1.1 mmol dicyclohexylcarbodiimide and continue stirring for another 6 hr at 4°. One mmol of 21 in 0.5 mL dry dimethylformamide, is added dropwise to the preceding reaction mixture and stirring continued under N 2 overnight at 4°. Remove the solvents in vacuo, solubilize the residue in acetonitrile and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 C0 3 containing 20% acetonitrile and eluted with a gradient of 0 to 1.0 M acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product (25). Dissolve 1 mmol of 25 in 10 mL 50% piperidine in DMF (v/v) . Remove the solvents in vacuo, solubilize the residue in 0.1 M NH 4 OH, and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 C0 3 and elute with a gradient to 1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product (26). To a stirred solution of 0.5 mmol of lytic peptide (Pep24) in 5 mL PBS, pH 7.4, at 4°, add 0.1 mmol of 23 in PBS dropwise. After 18 hr, chromatograph over a molecular sizing column to separate the product (27) from unreacted starting materials. c. trimeric fusogenic peptide covalently linked to a polycation through an acid sensitive, reducible spacer. tyr-lys-ala-(lys)^trp-lys-CON^ NHCO-C^-C^-CONH-CR j CR j -NH-R" (30) where R" = e-R' e-R' e-R' -β-C0-acoπityl-γ-C0HN-glu-ly Is-gly-lyIs-gly-pro-gly-gly-lyIs-α f γ-diaminobutyric-C0NH 2 with the γ-carboxyl of glu 1 in amide linkage with the γ-amino of α,γ- diaminobutyric acid 10 and R' = Pep24-SS-CH 2 CH 2 CO- The Series VI synthetic route (New compounds 28 -30)is shown in Figure 29. Detailed experimental procedures. Dissolve 1 mmol of 18 in 2 mL dry DMF, add 1.1 mmol N- hydroxysuccinimide and 1.1 mmol dicyclohexylcarbodiimide and continue stirring for another 6 hr at 4°. One mmol of 21 in 0.5 mL dry dimethylformamide, is added dropwise to the preceding reaction mixture and stirring continued under N 2 overnight at 4°. Remove the solvents in vacuo, solubilize the residue in acetonitrile and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 C0 3 containing 20% acetonitrile and eluted with a gradient of 0 to 1.0 M acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product (28). Dissolve 1 mmol of 28 in 10 mL 50% piperidine in DMF (v/v). Remove the solvents in vacuo, solubilize the residue in 0.1 M NH 4 OH, and apply to an anion exchange resin equilibrated in degassed 0.05 M NH 4 CO s and elute with a gradient to 1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product (29). To a stirred solution of 0.5 mmol of lytic peptide (Pep24) in 5 mL PBS, pH 7.4, at 4°, add 0.1 mmol of 29 in PBS dropwise. After 18 hr, chromatograph over a molecular sizing column to separate the product (30) from unreacted starting materials. Peptides such as Pep7 through PeplO are nuclear localization sequences which are used to target the inserted DNA to the nucleus. One skilled in the art recognizes that the peptides shown in Figure 18 are only examples of this class of peptides and that there are a wide variety of other nuclear localization sequence peptides which can be used. H 2 N-Tyr-e-N-lys-Pepll-CONH, and γ-N-[N-12-methoxy-6- chlorodiridinyl-HN-tyr-e-N-lys-Pepll-CO]-2-N-(2-methoxy-6- chloroacridinyl) diaminobutanoyl-COHN 2 were prepared by standard solid phase peptide synthesis. One skilled in the art recognizes that any amino acid polymer such as H 2 N-(arg-ala) n -COOH, histones and other DNA binding cationic polypeptides and proteins which form an α-helix, can be substituted for the lys-ala template. The -NH-(lys- ala)_.-CO unit can be extended. The useful range is from 2 to greater than 100 depending on the sequence of the inserted DNA, the target, uptake and specificity. The sequence position of the e-N-substituted- lys residue can be either amino-terminal or carboxyl-terminal. The substitution can be any amino reactive DNA binding dye as well as the acridine moiety. Examples of DNA binding dyes include thiaxanthenones, lucanthone, hycanthone, phenanthrenemethanol, metallointercalation reagents, tilorone, napthiophene, phenanthridiniums, dimidium, ethidium, propidium, quinacrine. In further embodiments, the spacers can be attached to the α- amino group of the N-terminal amino acid, and/or the carboxyl group of the C-terminal amino acid, rather than the e-amino group of lysine, to reduce immunological response to the ligand. C. In addition to the above components, it has also been found that fusion competent virus can be used to target the inserted DNA. Figure 4 shows the schematic procedure for preparing a fusion competent virus for use in the present invention. A variety of fusion competent virus can be used. As an example, adenovirus can be prepared in two separate ways. To a stirred solution of 10 mg of fusion competent adenovirus in 5 mL PBS, pH 7.4, at 4°, add 0.3 mL of 20 mM succinimidyl 3(2-pyridyldithio)propionate in ethanol dropwise. After 60 min, dialyze against 3 changes of 0.5 L PBS, pH 7.4, at 4°, each for 2 hr. Alternatively, to a stirred solution of 10 mg of fusion competent adenovirus in 5 mL PBS, pH 7.4, at 4°, add 0.3 mL of 20 mM 2- iminothiolane HCl in ethanol dropwise. After 60 min, dialyze against 3 changes of 0.5 L PBS, pH 7.4, at 4°, each for 2 hr. D. Recombinant Peptides 12, 13, and 16, 10 mg, prepared and purified by published methods (Bayne et al., J.Biol.Chem. 264:11004- 11008, 1988), are dissolved in 10 mL 50 mM NH 4 OH, pH 8.5. Aliquots are removed at 2 hr intervals to determine the extent of cyclization of N- terminal glutamine to pyroglutamate. When this reaction is complete, the solution is lypholyzed and then dissolved in 5 mL PBS, pH 7.4. The Peptides 12, 13, 16, and 17 are further reacted with either succinimyl 3(2-pyridyldithio) propionate or 2-iminothiolane as described for adenovirus (Example 1C) . E. Further, it has been found that either monoclonal or polyclonal IgG can be used to target the inserted DNA. Generally, the IgG is cleaved with immobilized pepsin to yield (Fab'-S-) 2 which is selectively reduced to Fab' -SH. Specifically, this includes: adding dropwise 0.5 mL 0.1 mM dithiothreitol to a stirred 5 mL solution of 1 nmol IgG F(ab') 2 at 4°, which was prepared by standard methods with immobilized pepsin. After 60 min, dialyze against 3 changes of 0.5 L PBS, pH 7.4, at 4°, each for 2 hr. F. Synthesis of component D. Standard continuous-flow solid phase synthetic methodologies are used to prepare H 2 N-his-leu-arg-arg- leu-arg-arg-arg-leu-leu-arg-glu-ala-glu-glu-gly-CONH g ,whichisreleased as a protected peptide, containing N™-Fmoc-his, N 9 -4-methoxy-2,3,6- trimethylphenylsulfonyl-arg, and glu-γ-Fmoc ester. Coupling of this protected peptide using DCCI with the appropriate protected peptide on the solid support, gives t-BOC-NH-CH-CONH-(CH 2 ) 5 COHN-Pepl-COO-resin CH 2 -CONH(CR,) 5 COHN-Pepl-CONH 2 . Deprotection of the asp-amino group, reaction with succinimidyl 3(2- pyridyldithio)propionate, deblocking and cleavage from the resin gives D. G. Synthesis of the asialoglycoprotein receptor ligand, component E. Dissolve 1 mmol N",N p -bis{hexanamido[ tris-(β- lactosylhydroxymethyl)methane]}aspartyl diamide (Meth. Enzymol. 138:424-429, 1987), in 20 mL PBS, pH 7.4, and combine with 1 mmol succinimyl 3(2-pyridyldithio) propionate, in 2 mL phosphate-buffered saline, pH 7.4. Dilute the reaction mixture 20-fold with water, apply to a cation exchange column to separate the desired product from unreacted starting material and other products, using a linear gradient formed from equal volumes of water and 2.0 M HCl. The appropriate fractions are pooled and lyophilized to obtain the product, E. H. Preparation of Compound I, L-tyrosyl-L-aspartoyl-_σis-{N-[6- [ [6-O-phosphory1-α-D-mannopyranosyl]oxy]hexyl]-L-alaninamide] is prepared as described (Carb. Res. 198:235-246 (1990), except that N-t- BOC-L-tyrosine is used in lieu of N-acetyl-L-tyrosine. Compound I is further reacted as described for Compound E. (Example 1G) . Example 2 Synthesis of Tetracationic DNA Binding Templates The overall schematic flow chart for the synthesis of these compounds is shown in Figure 5. The chemical pathway of synthesis is shown below. The Roman numerals are used to identify the specific compounds. Dissolve 2 mmol of free base l,4-diaminobutan-2-ol (II) (Meth. Enzymol. 94:431-433, 1983), in 5 mL of ethanol, add 4.1 mmol of acrylonitrile and allow to stand overnight at room temperature. Cool in an ice bath and saturate the solution with anhydrous NH 3 at 0°. Add about 5 mL of sponge nickel hydrogenation catalyst and shake under H 2 on a Paar low-pressure hydrogenator until the theoretical amount of H 2 is consumed. Remove the catalyst by filtration and wash the catalyst with ethanol. Combine filtrate and washings, then remove the ethanol in vacuo. Chromatograph on a cation exchange column to separate the desired product (R and S IV) from unreacted starting material and other products, using a linear gradient formed from equal volumes of water and 2.0 M HCl. Resolve the enantiomers of IV on a chiral column such as (R)-N-3,5-dinitrobenzoylleucine-silica (Baker) by a gradient of 2- propanol, from 0 to 20% in hexane. Br-CR j -CH-CR j CR j -Br (I) + NH 4 OH OH Ha -CHa-CH-CRjCRi-NR, (II) OH 4. CR j =CH-CN CN-(CR,) 2 - H-CH 2 -CH-( CH_) 2 ~NH-(CR-,) 2 -CN (III) OH 4- R j N-(CHa) 3 -NH-CH 2 -CH-(CR,) 2 -NH-(CR ; ) 3 -NH 2 (IV) OH Next combine 2 mmol (S)-hydroxyspermine (IV), 8 mmol 4- pyrrolidinopyridine and 8.2 mmol benzyloxycarbonyl anhydride (Z z O)in 40 mL anhydrous benzene and stir overnight at room temperature under N 2 . Separate the desired product (V) by solid phase extraction on phenyl- silica and elution with a linear gradient of ethyl acetate from 0 to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product as an amorphous solid. IV + Z 2 -0 4. Z-NH- ( CH 2 ) 3 -N-CH 2 -CH- ( CH 2 ) 2 -N- ( CH 2 ) 3 -NH-Z ( V ) OH + BrCR j COOCH g , NaOCH 3 4. Add 4 mL of dry benzene to 1 mmol I^COg and 2 mmol 18-crown-6 and stir for 20 min. Add 2 mmol V in 4 mL of dry benzene, followed by 2 mmol methyl bromoacetate in 2 mL benzene. After 4 hr, add 25 mL of water and extract with 3 portions of 25 mL benzene. Remove the solvent in vacuo and dissolve the residue (IV) in 10 mL ethanol containing 2 mmol potassium hydroxide. After overnight at room temperature, the solution is transferred to a separatory funnel, to which 2 mmol of HCl, 5 mL of water and 25 mL of benzene are added. After extraction with 3 additional portions of benzene, the combined organic phase is taken to dryness in vacuo, redissolved in a minimum volume of ethyl acetate, dried over 10% w/v anhydrous Na 2 S0 4 overnight. The organic phase is decanted and diluted with a sufficient amount of petroleum ether to create slight turbidity and cooled at 4° to promote crystallization of VII (1,4,9,12-tetrabenzyloxycarbonyl-l,12-diamino-6-carboxymethoxy-4,9- diazadodecane) . Z Z Z-NH- ( CH 2 ) 3 -N I -CH 2 -CH- ( CR; ) 2 -N I- ( CR; ) 3 -NH-Z ( VI ) OCR j COOCHg OH " Z-NH- ( CH 2 ) 3 -N-CH 2 -CH- ( CH 2 ) 2 -N- ( CH 2 ) 3 -NH-Z (VII ) OCR j COOH Dissolve 1 mmol of VII in 2 mL dry dimethylformamide, add 3.0 mmol l-ethyl-3-[3-(dimethylamino)propyl) ] carbodiimide, stir 2 hr, then add.1.1 mmol N-hydroxysuccinimide and stir for an additional 6 hr at room temperature. This solution is added dropwise to 1 mmol of A in 0.5 mL dry dimethylformamide. Stirring is continued in the dark under N 2 for an additional 4 hr. When the reaction is complete, as monitored by thin layer chromatography, 15 mL oxygen-free water is added to precipitate the product. The product is collected by centrifugation, washed and dissolved in oxygen-free 0.1 M NH 4 OH. The solution is applied to an anion exchange resin equilibrated in degassed 0.1 M NH 4 C0 3 containing 20% acetonitrile. The γ-isomer is separated from unreacted starting materials and the α-isomer by a gradient from 20% to 50% acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product. Alternative compounds in this series, for example Vlllb and VIIIc, are made by substituting the appropriate starting material containing biotin, lipoic acid or other substituent. VII + H^-C-^CR j -S-S-CR-CR j -NH-R Where R is A, B, G, P, J, Pep 12 through Pep 26 or H. 4- Z Z Z-NH-(CH 2 ) 3 -N i-CH 2 -CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NH-Z OCHaCO-rø-Cl^CH^S-S-CRjCRj-NH-R Where R is A (Villa), B (Vlllb) , G (VIIIc) or H (IX). Dissolve 1 mmol Villa or Vlllb or VIIIc or IX in 20 mL glacial acetic acid containing 30% HBr and stir overnight at room temperature in the dark under N 2 . Add 30 mL diethyl ether to precipitate the product. Wash the product until the odor of acetic acid is gone. Dissolve the solid in oxygen-free 0.1 M NH 4 OH and apply to an anion exchange resin equilibrated in degassed 0.1 M NH 4 C0 3 containing 20% acetonitrile. The product Xa or Xb or Xc or XI is separated from unreacted starting materials by a gradient of 0 to 90% acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product. Where R is A (Xa) , B (Xb , G (Xc) or H (XI). Dissolve 2 mmol XI purified by chromatography as was done for IV, in 10 mL oxygen-free 0.01 M NH 4 C0 3 containing 2 mmol dithiothreitol and stir for 2 hr. Bring the solution to pH 5 with IN HCl and apply the solution to a cation exchange resin equilibrated in degassed water. The product XII is isolated by a gradient from 0.0005 M to 2.0 M HCl. The appropriate fractions are pooled and lyophilized to obtain the product. This is followed by reactions described above to yield Xllld, Xllle and XHIf. XI + dithiothreitol 4. NH 2 -(CH 2 ) 3 -NH-CH 2 -CH-(CH 2 ) 2 -NH-(CH 2 ) 3 -NH 2 (XII) OCR-CONH-CR j CR j -SH A C 5 H 4 N-S-S-CH 2 CH 2 -CONH-R; Where R is D, E or F. NH 2 -(CH 2 ) 3 -NH-CH 2 -CH-(CH 2 ) 2 -NH-(CH 2 ) 3 -NH 2 OCH 2 CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-R Where R is D (XHId) , E (Xllle) or F (XHIf) . Dissolve 1 mmol XII in 2 mL phosphate-buffered saline, pH 7.4, and combine with 1 mmol of further reacted (see above) A, B or G in 2 mL phosphate-buffered saline, pH 7.4. Dilute the reaction mixture 20- fold with water, apply to an anion exchange resin equilibrated in degassed 0.1 M NH 4 CO s containing 20% acetonitrile. The product XIa, Xlb or XIc is separated from unreacted starting materials by chromatography in 0.1 M NH 4 C0 3 containing 20% acetonitrile. The appropriate fractions are pooled and lyophilized to obtain the product. XII + C g ^N-S-S-CR j CR j -NH-R Where R is A, B or G. 4- NH 2 -(CR ; ) g -NH-CR j -CH-(CR ; ) 2 -NH-(CR,) 3 -NR ; OCR-CONH-O^CR j -S-S-CR j CH g -NH-R Where R is A (Xia), B (Xib) or G (Xic). Dissolve 1 mmol of VII in 2 mL dry dimethylformamide, add 3.0 mmol l-ethyl-e-[3-(dimethylamino)propyl)carbodiimide and stir 2 hr, then add 1.1 mmol N-hydroxysuccinimide and continue stirring for another 6 hr at room temperature. This solution is added dropwise to 5 mmol of 1,6-diaminohexane in 20 mL dry dimethylformamide, and stirring continued for an additional 24 hr. Remove the solvent in vacuo . Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate from 0 to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product as an amorphous solid (XIV) . Next combine 1 mmol XIV in dry 10 mL benzene with 1.1 mmol succinimidyl 3(2- pyridylthio)propionate, stir for 2 hr at room temperature, and then remove the solvent I in vacuo . The resultant product is XV. VII + H 2 N-(CH 2 ) 6 -NH 2 4. - Z Z Z-NH-(CR,) 3 -N I-CH 2 -CH-(CR ; ) 2 ~N I-(CR ; ) 3 -NH-Z (XIV) OCHaCONH-C^C^C^C^C^CH g -N^ 4- succinimidyl 3(2-pyridyldithio)propionate Z Z Z-NH-(CH 2 ) 3 -N I-CH 2 -CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NH-Z (XV) OCH 2 CONH-(CR j ) 6 -NHCO-CH 2 CH 2 -S-S-C 5 H 4 N Dissolve 1 mmol XV, in 20 mL glacial acetic acid containing 30% Hbr and stir overnight at room temperature in the dark under N 2 . Add 30 mL diethyl ether to precipitate the product. Wash the product until the odor of acetic acid is gone. Dissolve the solid in oxygen-free 0.1 M NH 4 OH. The solution is applied to an anion exchange resin equilibrated in degassed 0.1 M NH 4 CO s containing 20% acetonitrile. The product is separated from unreacted starting materials by a gradient of 20 to 80% acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product: NH 2 -(CH 2 ) 3 -NH-CH 2 -CH-(CH 2 ) 2 -NH-(CH 2 ) 3 -NH 2 (XVI) OCH 2 CONH-(CH 2 ) 6 -NHCO-CH 2 CH 2 -S-S-C 5 H 4 N To a stirred solution of 10 mg of Fab' -SH in 5 mL PBS, pH 7.4, at 4°, add 0.3 mL of 10 Mm XVI in ethanol dropwise. After 60 min. dialyze against 3 changes of 0.5 L PBS, pH 7.4, at 4°, each for 2 hr. NR,-(CR;) 3 -NH-CH 2 -CH-(CR;) 2 -NH-(CH 2 ) 3 -NR, OCR j CONH-(CR ; ) 6 -NHCO-CR,CR ; -S-S-Fab' (XVIlg) Combine 2 mmol IX, 2 mmol 4-pyrrolidinopyridine and 3 mmol succinic anhydride in 40 mL anhydrous benzene and stir overnight at room temperature under N 2 . Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product as an amorphous solid. This is followed with DCCI, in situ coupling with resin bound protected peptide, Pep2-COOH, using standard solid phase synthetic techniques followed by deprotection and release from the resin to yield XlXh. IX + succinic anhydride Z-NH-(CH 2 ) 3 -N-CH 2 -CH-(CH 2 ) 2 -N-(CH 2 ) 3 -NH-Z (XVI11) OCH 2 CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -NHCO-CH 2 CH 2 -COOH NH 2 -(CH 2 ) 3 -NH-CH 2 -CH-(CH 2 ) 2 -NH-(CH 2 ) 3 -NH 2 (XlXh) OCH 2 CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -NHCO-CH 2 CH 2 -CONH-Pep2-COOH Dissolve 1 mmol of VII in 2 mL dry dimethylformamide, add 3.0 mmol l-ethyl-3-[3-(dimethylamino)propyl)carabodiimide and stir 2 hr, then add 1.1 mmol N-hydroxysuccinimide and continue stirring for another 6 hr at room temperature. This solution is added dropwise to 5 mmol of methyl 6-amonohexanoate in 20 mL dry dimethylformamide, and stirring continued for an additional 24 hr. Remove the solvent in vacuo . separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate from 0 to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product (XX) as an amorphous solid. VII + H^-(CH 2 ) 5 -COOCH 3 4. Z Z I I Z-NH- ( CH 2 ) 3 -N-CH 2 -CH- ( CH 2 ) 2 -N- ( CH 2 ) 3 ~NH- Z ( XX ) OCH 2 CONH- ( CH 2 ) 5 -COOCH 3 + OH- Dissolve 2 mmol XX in 10 mL ethanol containing 2 mmol potassium hydroxide. After overnight at room temperature, the solution is transferred to a separate runnel, to which 2 mmol of HCl, 5 mL of water and 25 mL of benzene is added. After extraction with 3 additional portions of benzene, the combined organic phase is taken to dryness in vacuo . Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate from 0 to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product (XXI) as an amorphous solid. Next, couple XXI to the amino terminal of the peptide on the support using standard solid phase peptide methods. z z Z-NH- ( CR,) g -N I-CR j -CH- ( CR, ) 2 -N I- ( CR, ) 3 -NH-Z (XXI ) OCR j CONH- ( CR 5 -COOH 1,4,9, 12-tetrabenzyloxycarbonyl-l , 12-diamino-6- [ ( 5 ' - carboxypenty 1 ) aminocarbonylmethoxy] -4, 9-diazadodecane 4- Ha- ( CHa) 3 -NH-CHa-CH- ( CR,) 2 -NH- (CR,) 3 -NH 2 OCR j CONH-(CHa) 5 -CONH-R-CONR j Where R is: Pep3 (XXIIi) , Pep4 (XXIIj), Pep5 (XXIIk) or Pep6 (XΣII1) Example 3 Further Tetracationic DNA Binding Templates The overall schematic flow chart of the synthesis of these compounds is shown in Figure 6. The chemical pathway of synthesis is shown below. A. After derivatization of the e-N-lys with succinic anhydride it is coupled to the ligands shown below: HaN-CHaCHa-S-S-CHaCRj- H-R Where R is A, B, β or H. deprotection and release from the resin yields H g N-tyr-lys-Pepll-CONR j I HN-CO(C-^)^COHN-C^CR j -S-S-CR j CR j -NH-R Where R is A (XXIIIa) , B (XXIIIb), G (XXIIIc) or H (XXIV). Following the procedures described for the synthesis of XII and XIII but substituting XXIV for XI yields XXV and XXVI. XXIV + dithiothreitol 4- HaN-tyr-lys-Pepll-CONR ; HN-CO(CH^-COHN-CR j CR j -SH (XXV) 4. C 5 H 4 N-S-S-CR,CR,-CONH-R; Where R is D r E or F. NH 2 -tyr-lys-Pebll-CONH 2 NHCO-CH 2 -CH 2 -CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-R Where R is D (XXVId) , E (XXVIe) or F (XXVIf) . After derivatization of the e-N-succinyl-lys with H 2 N-CH 2 CH 2 -S-S- CH 2 CH 2 -NH-t-BOC and deblocking, the ligand Pep2 is synthesized on the resin using standard solid phase techniques. Deblocking and cleavage from the resin yields XXVIIh. H 2 N-tyr-lys-Pepll-CONH 2 (XXVIIh) (CH 2 ) 4 NHCO-(CH 2 ) 2 -CONH-(CH 2 ) 2 -S-S-(CH 2 ) 2 -CONH-Pep2-COOH The resin bound lys-e-NH-CO(CH 2 ) 5 NH 2 intermediate was coupled with succinimidyl 3(2-pyridyldithio) ropionate and then deprotected and cleaved to yield XXVIII. H 2 N-tyr-lys-Pepll-CONH 2 (XXVIII) (CR,) 4 NHCO-(CR j ) 5 -NHCO-CR;CR ; -S-S-C 5 H 4 N To a stirred solution 10 mg of FAB' -SH in 5 mL PBS, pH 7.4, at 4°, add 0.3 mL of 10 mM XVI in PBS, pH 7.4, dropwise. After 60 min. dialyze against 3 changes of 0.5 L PBS, pH 7.4, at 4°, each for 2 hr. H 2 N-tyr-lys-Pepll-CONH 2 (CR ; ) 4 NHCO-(CH 2 ) 5 -NHCO-CR-CH 2 -S-S-Fab' (XXIXg) The nuclear localization sequence was added by standard solid phase synthetic methods to the lys-e-NH-CO(CH 2 ) 5 NH 2 intermediate of the resin bound protected peptide for carboxyl to amino orientation or to the N-succinyl derivative of e-N-lys for amino to carboxyl orientation. Deprotection and release from the resin yields: H 2 N-tyr-lys-Pepll-CONH 2 ( CR, ) 4 NHCO- ( CR ; ) j -NHCO-R-NR, Where R is Pep7 ( XXXi) , Pep8 ( XXXj ) , Pep9 (XXXk) or PeplO ( XXXI ) or R j N-tyr-lys-Pepll-CONR, ( CH 2 ) 4 NHCO- ( CR ; ) 2 -CONH-R-CONH 2 Where R is Pep3 (XXXIi), Pep4 (XXXIj), Pep5 (XXXIk) or Pep6 (XXXII) B. PARENT COMPOUND: N-(ligand moiety)-HN-tyr-lys-lys-ala-lys- ala-lys-ala-lys-CONH 2 , prepared by standard solid phase peptide synthesis. It is obvious that any amino acid polymer, such as H 2 N- (lys) n -COOH, R,N-(arg-ala) n -COOH, histones, and other DNA binding cationic polypeptides and proteins which form an α-helix, could be substituted for the lys-ala template. The -HN-(lys-ala) n -CO unit can be extended from 4 to more than 100. The sequence position of the residue bearing the spacer-ligand moiety can be either amino-terminal or carboxyl-terminal. In another embodiment, the ligand-spacer moiety is linked through a disulfide bond to cys as either the N-terminal or C-terminal residue. The resin bound protected peptide containing a deblocked α-amino- tyr moiety is the synthetic intermediate for preparation of templates for reductive release of a plasma membrane receptor ligand. After derivatization of the α-amino-tyr with succinic anhydride, the following ligands are coupled to the resin bound protected peptide. R J N-CR J CR.-S-S-CR J CR J -NH-R where R = γ-amide of the glutamyl moiety of folic acid (A) = biotin (B) = lipoic acid (G) -------- H Deprotection and release from the resin gives : tyr-lys-lys-ala- lys-ala- lys-ala-lys-CONHa I HN-CO ( CR j ) JJ -COHN-CR J CR J -S-S-CR J CR J -NH-R whereR = γ-amide of the glutamyl moiety of folic acid (XXIIIa) = biotin (XXIIIb) = lipoic acid (XXIIIc) = H Example 4 Synthesis of Hexacationic DNA Binding Template A schematic flow chart of the synthesis of these compounds is shown in Figure 7. The chemical pathway of synthesis is shown below. Dissolve 2 mmol of succinic monoamide in 2 mL dry DMF, add 4.0 mmol l-ethyl-3-[3-(dimethylamino)propyl)carbodiimide and stir 2 hr. Then add 2.1 mmol N-hydroxysuccinimide and continue stirring for another 6 hr at room temperature. Combine this in dropwise fashion to 1 mmol (S)-hydroxyspermine (IV) in 2 mL dry DMF. After stirring overnight at room temperature, remove the solvent in vacuo, dissolve in water, and apply the solution to a cation exchange resin. The product is isolated by a gradient to 2.0 M HCl. The appropriate fractions are pooled and lyophilized to obtain XXXII. Dissolve 2 mmol XXXII in 5 mL dry toluene and add 10 mmol sodium bis(2-methoxyethoxy)aluminum hydride in toluene in small aliquo s over 30 min. After 2 hr, add 10 mL ethyl acetate, then remove the solvents in vacuo. Dissolve the solids in water, adjust the pH to 3 with HCl and apply to a cation exchange resin. The product is isolated by a gradient to 2.0 M HCl. The appropriate fractions are pooled and lyophilized to obtain the product. H 2 N-(CH 2 ) 3 -NH-CH 2 -CH-(CH 2 ) 2 -NH-(CH 2 ) 3 -NH 2 (IV) OH + I^NCOCR j CR j COOH I HgNCOCR j CR j CONH- ( CH 2 ) 3 -NH-CH 2 -CH- ( CR; ) 2 -NH- ( CH 2 ) 3 -NHCOCH 2 CH 2 CONH 2 I (XXXII ) OH 4. Red-Al HgN ( C ; ) 4 -NH- ( CH 2 ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CH 2 ) 3 -NH- ( CH 2 ) 4 -NH 2 I (XXXIII) OH It is obvious to one skilled in the art that homologs with additional - (CH 2 ) 4 -NH 2 , or -(CH^g-NH g units can be made by repeating the reactions using XXXIII in lieu of IV with H 2 NCOCH 2 CH 2 COOH or CH 2 =CHCN. In reactions similar to the above reaction where IV is converted to VII, XXXIII can be converted to XXVI. XXXIII + Z 2 0 4- Z Z Z Z Z-HN-(CR ; ) 4 -N I-(CR ; ) 3 -NI-CH 2 -CH-(CR ; ) 2 -N I-(CR ; ) 3 -NI-(CH 2 ) 4 -NH-Z I (XXXIV) OH 4. BrCH 2 -COOCH 3 , NaOCH 3 Z Z Z z Z-HN- ( CR; ) 4 -N I- ( CH 2 ) 3 -NI-CR,-CH- ( CR, ) 2 -N I- ( CH 2 ) 3 -NI- ( CH 2 ) 4 -NH- Z I (XXXV) OCR.COOCH 3 + OH " 4- Z Z Z Z Z-HN-(CH 2 ) 4 -N I-(CH 2 ) 3 -NI-CH 2 -CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NI-(CH 2 ) 4 -NH-Z I (XXXVI) OCR,COOH 1,5,9, 14, 18, 22-hexabenzyloxycarbonyl-l, 22-diamino-ll-carboxymethoxy- 5 , 9 , 14, 18-tetraazadocosane In reactions similar to the above reactions where VII is converted to the VII and XI series, XXXVI can be converted to the XXXVII and XXXIX series. XXXVI + HJJN-CRJCHJJ-S-S-CRJCRJ-NH-R Where R is A, B, G or H. I Z Z Z Z Z-HN-(CH 2 ) 4 -N I-(CH 2 ) 3 -NI-CH 2 -CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NI-(CH 2 ) 4 -NH-Z OC^CONH-CRJCRJ-S-S-CRJCRJ-NH-R Where R is A (XXXVIIa) , B (XXXVIIb), G (XXXVIIc) or H (XXXVIII). 4- HBr, CH 3 COOH HaN-(CH 2 ) 4 -NH-(CR,) 3 -NH-CH 2 -CH-(CH 2 ) 2 -NH-(CH 2 ) 3 -NH-(CH 2 ) 4 -NH 2 OCRJCONH-CRJCRJ-S -S -CRJCHJ-NH-R Where R is A (XXΣIXa), B (XXΣIXb), G (XXXIXc) or H (XL). In reactions similar to the above for the conversion of XI to the XIII series XL is converted to the XLII series. XL + dithiothreitol I H 2 N-(CHa) 4 -NH-(CH 2 ) 3 -NH-CH 2 -CH-(CH 2 ) 2 -NH-(CH 2 ) 3 -NH-(CHa) 4 -NH 2 (XLI) OCH 2 CONH-CR.CH 2 -SH 4, CgR j N-S-S-CR j CR j -CONH-R; Where R is D, E or F. NH 2 - ( CH 2 ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CH 2 ) 3 -NH 2 OCR-CONH-CR j CR j -S-S-CR j CR j -CONH-R Where R is D (XLIId), E (XLIIe) or F (XLIIf) In reactions similar to the above conversion of XII to the XI series, XLI is converted to the XXXIX series. XLI + C 5 H 4 N-S-S-CH 2 CH 2 -NH-R Where R is A, B or G. 4. H 2 N- ( CH 2 ) 4 -NH- ( CR, ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CH 2 ) 3 -NH- ( CH 2 ) 4 -NH 2 OCH 2 CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -NH-R Where R is A (XXXIXa) , B (XXXIXb) or G (XXXIXc) . In reactions similar to the above conversion of VII to XIV and XVIIg, XXXVI is converted to XLIV and XLV g. XXXVI + H^N-(CH 2 ) 6 -NH 2 4. Z Z Z Z Z-HN-(CH 2 ) 4 -N I-(CH 2 ) 3 -NI-CH 2 -CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NI-(CH 2 ) 4 -NH-Z (XLIII) OCH j CONH-C^C^CH^R-CR-CR j -NR; 4. + succinimidyl 3 (2-ρyridyldithio) propionate Z Z Z Z | | | | Z-HN- (CR,) 4 -N- (CH 2 ) 3 -N-CH 2 -CH- (CR,) 2 -N- (CH 2 ) 3 -N- (CR;) 4 -NH-Z (XLIV) OCR j CONH- ( CR j ) 6 -NHCO-CR j CH 2 -S-S-C 5 H 4 N 4. + HBr, CH j COOH H g N- ( CHg ) 4 -NH- ( CH 2 ) 3 -NH-CH 2 -CH- ( CR; ) 2 -NH- ( CR j ) 3 -NH- ( CH 2 ) 4 -NH 2 I (XL ) OCR j CONH-(CR,) β -NHCO-CH 2 CH 2 -S-S-C 3 H 4 N 4. + Fab'-SH R j N-(CH 2 ) 4 -NH-(CR j ) 3 -NH-CH 2 -CH-(CH 2 ) 2 ~NH-(CH 2 ) 3 -NH-(CR;) 4 -NH 2 I (XLVIg) OCR j CONH-(CR j )e-NHCO-CR-CR j -S-S-Fab' In reactions similar to the above conversion of IX to X Xh, XXXVIII is converted to XLVIIIh. XXXVIII + succinic anhydride 4- Z Z Z Z Z-HN-(CH 2 ) 4 -N I-(CH 2 ) 3 -NI-CH 2 -CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NI-(CH 2 ) 4 -NH-Z (XLVII) OCRJCONH-CRJCHJ-S HOOC-CH 2 CR,-COHN-CH 2 CRJ-S ^N-(CH 2 ) 4 -NH-(CH 2 ) 3 -NH-CH 2 -CH-(CH 2 ) 2 -NH-(CH 2 ) 3 -NH-(CHa) 4 -NH 2 OCH 2 CONH-(CH 2 ) 2 S (XLVIIIh) HOOC-Pep2-HNOC-CH 2 CH 2 -COHN-CH 2 CH 2 -S In reactions similar to the above conversion of VII to XXII series, XXXVI is converted to the LI series. XXXVI + H 2 N-(CH 2 ) 5 -COOCH 3 4- Z-HN- ( CR j ) Z Z Z Z Z-HN-(CH 2 ) 4 -N I-(CH 2 ) 3 -NI-CH 2 -CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NI-(CH 2 ) 4 -NH-Z (L) OCH 2 CONH- ( CR j ) 5 -COOH 1, 5,9 , 14, 18,22-hexabenzyloxycarbonyl-l,22-diamino-ll-[N(5 ' - carboxypentyl ) carbonylmethoxy-5 , 9 , 14, 18-tetraazadocosane Ha- (CR j ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CR j ) 3 -NH2 OCR j CONH-(CH 2 ) g -CONH-R-CONR ; Where R is: Pep3 (Lli) , Peρ4 (LIj) , Pep5 (LIk) or Pep6 (LI1) Example 5 Intercalating hexacationic DNA Binding Templates A schematic flow chart for the. synthesis of these compounds is shown in Figure 8. The chemical pathway of synthesis is shown below. Combine 2 mmol III (resolved S enantiomer) , 4 mmol 4-pyrrolidinopyridine and 4.1 mmol benzyloxycarbonyl anhydride in 40 mL anhydrous benzene and stir overnight at room temperature under N 2 . Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product (LII) as an amorphous solid. Ill (resolved S enantiomer) + t-BOC 2 0 4. t-BOC t-BOC CN-(CR j ) 2 -N I-CH 2 -CH-(CR,) 2 -N I-( CΑ,) 2 -CN (LII) OH + BrCH 2 -COOCH 3 , NaOCH 3 Add 4 mL of dry benzene to 1 mmol K g CO g and 2 mmol 18-crown-6 and stir for 20 min. Add 2 mmol LIII in 4 mL of dry benzene, followed by 2 mmol methyl bromoacetate in 2 mL benzene. After 4 hr., add 25 mL of water and extract with 3 portions of 25 mL benzene. Remove the solvent in vacuo to obtain the product: t-BOC t-BOC CN- ( CR; ) 2 - IN-CR,-CH- ( CR ; ) 2 -N I - ( CR, ) 2 -CN ( LI 11 ) OCH2-COOCH 3 4. H 2 , Raney Ni t-BOC t-BOC H 2 N- ( CH 2 ) 3 -N I-CR,-CH- ( CH, ) 2 -N I- ( CR, ) 3 -NH 2 ( LI V ) OCR j -COOCR j Dissolve 5 mmol LIV in 10 mL dry pyridine containing 0.1 mmol dimethylaminopyridine and 15 mmol triethylamine. Add dropwise 11 mmol 6,9-dichloro-2-methoxyacridine on any DNA binding dye that reacts specifically with amino group in 10 mL dry pyridine to the stirred solution. Stir for over 1 hr at room temperature. The solvents are removed in vacuo, and the mixture is redissolved in acetonitrile for solid phase extraction on phenyl-silica and elution with a linear gradient of acetonitrile to 50% in hexane. The appropriate fractions were pooled and the solvent evaporated to obtain the product as an amorphous solid. t-BOC t-BOC C 14 H 9 ClNO-HN- ( CH 2 ) 3 -N I-CR j -CH- ( CH j , ) 2 -N I- ( CH 2 ) 3 -NH-C 14 H g ClNO ( V ) I OCR j -COOCR, + OH _ Dissolve 2 mmol LV in 10 mL ethanol containing 2 mmol potassium hydroxide. After overnight at room temperature, the solution is transferred to a separatory funnel, to which 2 mmol of HCl, 5 mL of water and 25 mL of benzene is added. After extraction with 3 additional portions of benzene, the combined organic phase is taken to dryness in vacuo. Dissolve in dry DMF for standard solid phase peptide synthesis-. t-BOC t-BOC C 14 H 9 ClNO-HN-(CH 2 ) 3 -N I-CH 2 -CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NH-C 14 H g ClNO (LVI) OCH 2 -COOH l,12-N,N'-di(2-me hoxy-6-chloroacridin l) amino-4,9-di-t- butyoxycarbonyl-6-carboxymethoxy-4,9-diaza-dodecane In reactions similar to the conversions in Example 2, the following products are obtained. One skilled in the art will recognize that the reaction conditions are similar, but that the starting material and end product will be different. LVI + H^-CH^R J -S-S-CR J CR J -NH-R I Where R is A, B, G or H. C 14 H g ClNO-HN- Where R is A (LVIIa) , B (LVIIb ) , G (LVIIc ) or H (LVIII ) . 4- HBr, CH g COOH C 14 H g ClNO-NH- (CR,) g-NH-CR j -CH- (CR;) 2 -NH- (CR j ) 3 -NH-C 14 HgClNO OC^CONH-CR j CR j -S-S-CI^CR j -NH-R Where R is A (LIXa ) , B (LIXb ) , G (LIXc) or H (LX ) . (b ) LX + dithiothreitol 4. C 14 H g ClNO-NH- ( CR j ) 3 -NH-CR,-CH- ( CR j ) 2 -NH- ( CR- ) 3 -NH-C 14 R,ClNO (LXI ) I OCRJCONH-CRJCHJJ-SH 1 Cg^N-S-S-CI^CR j -CONH-R; Where R is D , E or F . C 14 H 9 ClNO-NH- ( CH 2 ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CH 2 ) 3 -NH-C 14 H 9 ClNO OCH2CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-R Where R is D (LXIId ) , E (LXIIe ) or F (LXIIf ) . LXI + CgH 4 N-S-S-CR j CH 2 -NH-R; Where R is A, B or G. 4. C 14 H g ClNO-NH- ( CH 2 ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 ~NH- ( CH 2 ) 3 -NH-C 14 H 9 ClNO OCH 2 CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -NH-R Where R is A (LIXa ) B (LIXb ) or G (LIXc ) . LVI + R j N- f CR j J g- R j 4. t-BOC t-BOC C 14 H θ ClNO-HN- ( CR, ) 3 -N-CH 2 -CH- ( CH 2 ) 2 -N- ( CH 2 ) 3 -NH-C 14 H β ClNO OCH 2 CONH- ( CH 2 ) 6 -NH 2 ( LXI 11 ) 4- succinimidyl 3 ( 2-pyridyldithio ) propionate t-BOC t-BOC C 14 HgClNO-HN- ( CH 2 ) 3 -N I-CH 2 -CH- ( CH 2 ) 2 - IN- ( CH 2 ) 3 -NH-C 14 H 9 ClNO ( LXIV ) OCH 2 CONH- ( CR j ) 6 -NHCO-CH 2 CH 2 -S-S-C 8 H 4 N 4. + HBr, CH g COOH C 14 H 9 ClNO-NH- ( CH 2 ) 3 -NH-CH 2 -CH- ( CR j ) 2 "NH- ( CH 2 ) 3 -NH-C 14 HgClNO ( LXV ) OCR j CONH- ( CH 2 ) g-NHCO-CHgC^-S-S-C 8 ^N 4. + Fab ' -SH C 14 H 9 ClNO-NH- ( CH 2 ) 3 -NH-CH 2 -CH- ( CH j ) 2 -NH- ( CR; ) 3 -NH-C 14 H 9 ClNO OCH2CONH- ( CR j ) 6 -NHCO-CR;CH 2 -S-S-Fab ' (LXVIg ) LVI + succinic anhydride t-BOC t-BOC C 14 H 9 ClNO-HN- ( CR ; ) 3 -N I-CR,-CH- (CH 2 ) 2 ~ IN- ( CR, ) 3 -NH-C 14 H 9 ClNO (LXVII ) OCH 2 CONH-CH 2 CH 2 -S HOOC-CH 2 CH 2 -OCHN-CH 2 CH 2 -S 4- C 14 H g ClN0-NH- ( CH 2 ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CH 2 ) 3 -NH-C 14 H 9 ClNO ( LXVI I Ih ) O-CR j -CONH- ( CH g ) 2 -S HOOC-Pep2 -HNOC-CH 2 CH 2 -HNOC-CH 2 CH 2 -S LVI + H 2 N(CR j ) 5 -COOCH 3 1 t-BOC t-BOC C 14 H 9 C1N0-HN- ( CR j ) 3 -N I-CH 2 -CH- ( CR ; ) 2 - IN- ( CH 2 ) 3 -NH-C 14 H 9 ClNO (LXIX) OCH 2 -CONH ( CR; ) 5 -COOCH 3 OH " t-BOC t-BOC C 14 H g ClNO-HN- ( CHa ) 3 -N I-CR,-CH- ( CR j ) 2 - IN- ( CR ; ) 3 -NH-C 14 H g ClNO (LXX) OCH 2 -CONH ( CR; ) g -COOH 1 , 12-N, N ' -di ( 2-methoxy-6-chloroacridinyl ) amino -4 , 9-di-t- butyoxycarbonyl-6- [N ( 5 ' -carboxypentyl ) aminocarbonylmethoxy ] -4 , 9-diaza- dodecane C 14 H 9 ClNO-NH- ( CHa ) 3 -NH-CR,-CH- ( CH 2 ) 2 -NH- ( CR, ) 3 -NH-C 14 H g ClNO OCH 2 CONH- ( CR, ) g-C0NH-R-C0NH 2 Where R is Pep3 (LXXIi), Pep4 (LXXIj), Pep5 (LXXIk) or Pep6 (LXXI1) Example 6 Further Intercalating hexacationic DNA Binding Template A schematic flow chart for the synthesis of these compounds is shown in Fig. 9. The chemical pathway of synthesis is shown below. In reactions similar to the conversions in Example 3, the following products are obtained. One skilled in the art will recognize that the reaction conditions are similar, but that the starting material and end products will be different. After derivatization of the e-N-lys with succinic anhydride, the following ligands are coupled to the resin bound protected peptide. R,N-CH 2 CH 2 -S-S-CH 2 CH 2 -NH-R Where R is A, B, G or H. deprotection and release from the resin yields: CONH, C 14 H 9 ClNO-NH-tyr-lys-Pepll-CONH-CH 2 CH 2 -C IH-NH-C 14 H 9 ClNO HN-CO ( CR, ) 2 -COHN-CH 2 CH 2 -S-S-CH 2 CH 2 -NH-R Where R is A (LXXIIa ) , B (LXXIIb ) , G (LXXIIc ) or H (LXXIII ) . LXXIII + dithiothreitol 4. CONH, C 14 HgClNO-NH-tyr-lys-Pepll-CONH-CH 2 CH 2 -C IH-NH-C 14 H 9 ClNO HN-CO ( CH 2 ) 2 -COHN-CR,CR j -SH (LXXIV) Where R is D (LXXVd ) , E (LXXVe ) or F (LXXVf ) . CONH 2 I C 14 H 9 ClNO-NH-tyr-lys-Pepll-CON H-CH 2 CH 2 .CH-NH-C 14 H 9 ClNO (LXXVIh ) ( CR, ) 4 -HNCO- ( CR, ) 2 CONH- ( CH 2 ) 2 -S HOOC-Pep2-HNOC-CR;CR;-S CONH 2 C 14 H 9 ClNO-NH-tyr-lys-Pepll-CONH-CH2CH 2 -CH-NH-C 14 H g ClNO ( CH 2 ) 4 NHCO- ( CH 2 ) 5 -NHCO-CH 2 CH 2 -S-S-C 5 H 4 N (LXXVII ) 4- + Fab ' -SH (LXXVIIIg ) The addition of the nuclear localization sequence yields: CONR, C 14 H g ClNO-NH-tyr-lys-Pepll-CONH-CHaCH 2 -CH-NH-C 14 H 9 ClNO ( C , ) 4 NHCO- ( CHa) 5 -NHCO-R-NR j where R is Pep7 (LXXIXi ) , Pep8 (LXXIX j ) , Pep9 , (LXXIXk) or PeplO (LXXIX1) . CO H 2 C 14 H g ClNO-NH-tyr-lys-Pepll-CONH-C Cl^-CH-NH-C 14 H g ClNO ( CH 2 ) 4 NHCO- ( CHa ) 2 -C0NH-R-C0NR; where R is Pep3 (LXXXi ) , Pep4 (LXXXj ) , Pep5 (LXXXk) or Pep6 (LXXX1 ) . Example 7 Dimeric Octacationic DNA Binding Templates A schematic flow chart for the synthesis of these compounds is shown in Figure 10. The chemical pathway of synthesis is shown below. Combine 2 mmol LIV, 1 mmol 4-pyrrolidinopyridine and 1.0 mmol benzyloxycarbonyl anhydride in 40 mL anhydrous benzene and stir overnight at room temperature under N 2 . Separate the product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate, 0 to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the produce (LXXXI) as an amorphous solid. LIV + 0.5 mole t-BOC 2 0 4. t-BOC t-BOC t-BOC-HN-(CH 2 ) 3 -N I-CR j -CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NH 2 (LXXXI) OCR j -COOCR, Combine 2 mmol LXXXI, 2 mmol 4-pyrrolidinopyridine and 3 mmol bis-(3-carboxyethyl)dithiol in 40 mL anhydrous benzene and stir overnight at room temperature under N 2 . Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product (LXXXII) as an amorphous solid. t-BOC -BOC t-BOC-NH-(CH 2 ) 3 -N I-CH 2 CH-(CH 2 ) 2 -N I-(CH 2 ) 3 -NH-CO-CH 2 CH 2 -SS-CH 2 CH 2 -C001-JLXXXII) OCH 2 -COOCH 3 Dissolve 2 mmol LXXXII in 2 mL dry DMF, add 4.0 mmol l-ethyl-3- [3-(dimethyl-amino)propyl)carbodiimide and stir 2 hr, then add 2.1 mmol N-hydroxysuccinimide and continue stirring for another 6 hr at room temperature, then combine with 2 mmol 3-[ (3' '-N-t-BOC-aminopropyl)-4'- N-t-BOC-aminobutyl]-N-t-BOC-aminopropylamine in 21 mL dry DMF. After stirring overnight at room temperature, remove the solvent in vacuo, separate the product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate, 0 to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product (LXXXIII) as an amorphous solid. t-BOC t-BOC t-BOC HN-(CR,),-N-CH j -CH-(CR,) 2 -N-(CH,) 3 -NHCO-CR,CR, OCH 2 -COOCH 3 S (LXXXIII) S I HN- ( CH 2 ) 3 -N- ( CHa ) 4 "N" ( CH, ) 3 -NHOC-CR;CH 2 t-BOC t-BOC t-BOC Dissolve 2 mmol LXXXIII in 10 mL ethanol containing 2 mmol potassium hydroxide. After overnight at room temperature, the solution is transferred to a separatory funnel, to which 2 mmol of HCl, 5 mL of water and 25 mL of benzene is added. After extraction with 3 additional portions of benzene, the combined organic phase is taken to dryness in vacuo. Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product (LXXXIV) as an amorphous solid. t-BOC t-BOC t-BOC HN I-(CH 2 ) 3 -N I-CR ; -CH-(CH 2 ) 2 -N I-(CR,) 3 -NHCO-CR,CH 2 OCH 2 -COOH S (LXXXIV) S HN-(CHa) 3 - -(CH 2 ) 4 -N-(CR,) 3 -NHOC-CR j CR j t-BOC t-BOC t-BOC Dissolve 1 mmol of LXXXIV in 2 mL dry dimethylformamide, add 3.0 mmol l-ethyl-3-[3-(dimethylamino)propyl)carbodiimide and stir 2 hrs., then add 1.1 mmol n-hydroxysuccinimide and continue stirring for another 6 hrs. at room temperature. This solution is added dropwise to 3 mmol of Ha -CHaCHa-NH-R (where R = A, B, G or H) in 20 mL dry dimethylformamide, and stirring continued for an additional 24 hrs. Remove the solvent in vacuo. Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate from 0 to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product as an amorphous solid. LXXXIV + HaN-CR j CHa-S-S-CR j CR j -NH-R where R is A, B, G or H. 4- t-BOC t-BOC t-BOC HN I-(CHa) 3 -N I-CH 2 -CH-( CH 2 > 2 "N I"(-* )s-NHCO-C^CR, OCR^-CONH-CRJCRJ-S S R-HN-CR,CR,-S S HN-(CR,) 3 -N-(CHa) 4 -N-(CR;)g-NHOC-CI^CR, t-BOC t-BOC t-BOC where R is A (LXXXVa), B (LXXXVb), G (LXXXVc) or H (LXXXVI) . Dissolve LXXXVa, 1 mmol, in 29 mL glacial acetic acid containing 30% HBr and stir overnight at room temperature in the dark under N 2 . Add 30 mL diethyl ether to precipitate the product. Wash the product until the odor of acetic acid is gone. Dissolve the solid in oxygen-free 0.1 M NH 4 OH. The solution is applied to an anion exchange resin equilibrated in degassed 0.1 M NH 4 C0 3 containing 20% acetonitrile. The product is separated from unreacted starting materials by a gradient of 20 to 80% acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product: H 2 N- where R is A (LXΣXVIIa), B (LΣΣXVIIb), G (LXXXVIIc) or H (LXΣΣVIII). Use the same procedures as for LΣXXVa-LΣΣΣVc, except substitute S-t-BOC-mercaptoethylamine for H 2 N-CH 2 CH 2 -SS-CH 2 CH 2 -NH-R. Then use the same procedures as for LΣXXVIIa-LΣΣΣVIIc. LXXΣIV + t-BOC-S-CR j CR j -NR, 4. t-BOC t-BOC t-BOC HN I- ( CR j ) 3 -N I -CH 2 -CH- ( CR; ) 2 -N I- ( CH 2 ) 3 -NHCO-CH 2 CH 2 OCH,-CONH-CH,CH -S S (LΣΣΣIX ) t-B IOC S I HN- ( CH 2 ) 3 -N- ( CH 2 ) 4 -N- ( CR, ) 3 -NHOC-CH 2 C IH 2 I I I t-BOC t-BOC t-BOC 4- HBr , CH g COOH H 2 N- ( CH 2 ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CR, ) 3 -NHCO-CH 2 CH 2 OCH 2 -CONH-CH 2 CH 2 -SH S ( XC ) S H 2 N- ( CH 2 ) 3 -NH- ( CH 2 ) 4 -NH- ( CH 2 ) 3 -NHOC-CH 2 ICH 2 4. C 5 H 4 N-S-S-CR j CR j -CONH-R; where R is D, E or F . H 2 N- ( CH 2 ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CH 2 ) 3 -NHCO-CH 2 CH 2 OCH 2 -CONH-CH 2 CH 2 -S S R-HNOC-CR,CR,-S S H 2 N- ( CH 2 ) 3 -NH- ( CH 2 ) 4 -NH- ( CH 2 ) 3 -NHOC-CH 2 CH 2 where R is D ( XCId ) , E ( XCIe ) or F ( XCIf ) Dissolve 1 mmol xc in 2 mL phosphate-buffered saline, pH 7.4, and combine with 1 mmol A', also dissolved in 2 mL phosphate-buffered saline, pH 7.4. Dilute the reaction mixture 20-fold with water, apply to an anion exchange resin equilibrated in degassed 0.1 M NH 4 C0 3 containing 20% acetonitrile. The product XIa is separated from unreacted starting materials by a gradient of 20 to 80% acetonitrile in 0.1 M NH 4 C0 3 . The appropriate actions are pooled by lyophilized to obtain the product. XC + CgH 4 N-S-S-CR ; CR,-NH-R; where R is A, B or G. 4- R,N- ( CHa ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CR, ) 3 -NHC0-CH 2 CH 2 OCH 2 -CONH-CR j CR j -S S R-HN-CR j CR j -S S H 2 N- ( CHa ) 3 -NH- ( CR, ) 4 -NH- (CH 2 ) 3 -NHOC-CR,CH 2 where R is A (LΣΣΣVIIa) , B (LΣΣΣVIIb) or G (LΣΣXVIIc) . Dissolve 1 mmol of LXΣΣIV in 2 mL dry dimethylformamide, add 3.0 mmol l-ethyl-3-[3-(diamethylamino)propyl) carbodiimide and stir 2 hr, then add 1.1 mmol N-hydroxysuccinimide and continue stirring for another 6 hr at room temperature. This solution is added dropwise to 5 mmol of 1,6-diaminohexane in 20 mL dry diamethylformamide, and stirring continued for an additional 24 hr. Remove the solvent in vacuo. Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate from 0 to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product as an amorphous solid. LΣXΣIV + I t-BOC t-BOC t-BOC I I I HN- ( CH 2 ) 3 -N-CH 2 -CH- ( CR, ) 2 -N- ( CH 2 ) 3 -NHCO-CR,CR j OCR j -CONH- ( CR; ) 6 -NR, S (XCII ) S ! HN- ( CH2 ) 3 -N- ( CR; ) 4 -N- ( CR, ) g-NHOC-CHaCR, t-BOC t-BOC t-BOC Combine 1 mmol XCII in dry 10 mL benzene with 1.1 mmol succinim dyl 3(2-pyridylthio) propionate, stir for 2 hr at room temperature, and then remove the solvent in vacuo. Dissolve XCIII, 1 mmol, in 20 mL glacial acetic acid containing 30% HBr and stir overnight at room temperature in the dark under N 2 . Add 30 mL diethyl ether to precipitate the product. Wash the product until the odor of acetate acid is gone. Dissolve the solid in oxygen- free 0.1 M NH 4 C0 3 . The solution is applied to an anion exchange resin equilibrated in degassed 0.1 M NH 4 C0 3 containing 20% acetonitrile. The product is separated from unreacted starting materials by a gradient of 20 to 80% acetonitrile in 0.1 M NH 4 C0 3 . The appropriate fractions are pooled and lyophilized to obtain the product. To a stirred solution of 10 mg of Fab'-SH in 5 mL PBS, pH 7.4, at 4°, add 0.3 mL of 10 mM ΣCIV in PBS, pH 7.4, dropwise. After 60 min, dialyze against 3 changes of 0.5 L PBS, pH 7.4, at 4°, each for 2 hr. Combine 2 mmol LXXΣVI, 2 mmol 4-pyrrolidinopyridine and 3 mmol succinic anhydride in 40 mL anhydrous benzene and stir overnight at room temperature under N 2 . Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product (ΣCVI) as an amorphous solid. t-BOC t-BOC t-BOC HN I- ( CR j ) 3 -N I -CH 2 -CH-CH 2 -CH 2 -N I-CH 2 -CH 2 -CH 2 -NHCO-CH 2 -CH 2 OCR,-CONH- ( CH 2 ) 2 -S S (ΣCVI ) HOOC-CHs-CR j -OCHN-CHaCH g -S S HN- ( CH 2 ) 3 -N- ( CH 2 ) 4 -N- (CH 2 ) 3 -NHOC-CH 2 CH 2 t-BOC t-BOC t-BOC Couple XCVI to the amino terminal of the peptide on the support using standard solid phase peptide methods, cleave from the resin and deprotect, and purify by ion exchange chromatography. H 2 N-(CR,) 3 -NH-CH 2 -CH-CH 2 -CH 2 -NH-CH 2 -CH 2 -CH 2 -NHCO-CR j -CH 2 (ΣCVIIh) OCR,-CONH-(CR j ) 2 -S S HOOC-Pep2-HNOC-CH 2 CH 2 -OCHN-CH 2 CH 2 -S S j N-(CH 2 ) 3 - H-(CH 2 ) 4 -NH-(CH 2 ) 3 -NHOC-CH 2 -CH 2 Dissolve 2 mmol LΣΣΣIV in 2 mL dry DMF, add 4.0 mmol l-ethyl-3- [3-(dimethylamino)propyl)carbodiimide and stir 2 hr, then add 2.1 mmol N-hydroxysuccinimide and continue stirring for another 6 hr at room temperature, then combine with 2 mmol methyl 6-aminohexanoate in 2 mL dry DMF. After stirring overnight at room temperature, remove the solvent in vacuo, separate the product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate, 0 to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product as an amorphous solid. Dissolve 2 mmol XCVI11 in 10 mL ethanol containing 2 mmol potassium hydroxide. After overnight at room temperature, the solution is transferred to a separatory funnel, to which 2 mmol of HCl, 5 mL of water and 25 mL of benzene is added. After extraction with 3 additional portions of benzene, the combined organic phase is taken to dryness in vacuo . Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate to 100% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product as an amorphous solid. t-BOC t-BOC t-BOC HN I- ( CH 2 ) 3 -N I -CH 2 -CH-CH 2 -CH 2 -N I-CH 2 -CH 2 -CH 2 -NHCO-CH 2 -CR j OCH 2 -CONH- ( CH 2 ) g-COOH S (ΣCIΣ ) S HN-(CR,) 3 -N-(CH 2 ) 4 -N-(CH 2 ) 3 -NHOC-CH 2 C IH 2 t-BOC t-BOC t-BOC Couple ΣC X to the amino terminal of the appropriate peptide on the support using standard solid phase peptide methods, cleave from the resin and deprotect, and purify by ion exchange chromatography. H 2 N- ( CR, ) 3 -NH-CH 2 -CH-CH 2 -CH 2 -NH-CH 2 -CH 2 -CH 2 -NHCO-CH 2 -CH 2 I I OCR j -CONH- ( CR j ) 5 -CONH-R-CONH 2 S S H 2 N—(CR,) 3 -NH-(CR,) 4 -NH-(CR,) 3 -NHOC-CH 2 C IH 2 where R is Pep3 (XCXi), Pep4 (XCXj), Pep5 (ΣCΣk) or Peρ6 (ΣCXl). Example 8 Further Dimeric Octacationic DNA Binding Templates A schematic flow chart for the synthesis of these compounds is shown in Figure 11. The chemical pathways of synthesis are shown below. After derivatization of the e-N-Lys or α-N-Tyr with succinic anhydride the following ligands are coupled to the resin bound protective peptide: H 2 N-CR j CR j -S-S-CH 2 CH 2 -NH-R where R is A, B or G. Deprotection and release from the resin yields : H 2 N-tyr-lys-Pepll-CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-Pepll-CONH 2 HN-CO- ( CH 2 ) 2 -COHN-CH 2 CH 2 -S-S-CH 2 CH 2 -NH-R where R is A ( Cla ) , B ( Clb ) or G ( CIc ) . After derivatization of the e-N Lys with t-BOC-S- (CH 2 ) 2 -COOH, deprotection and release from the support yields : HaN-tyr-lys-Pepll-CONH-CHaCHa-S-S-CHaCHa-CONH-Pepll-CONR j HN-CO-CR j CR,-SH ( CII ) To a stirred solution of 10 mg of D or E in 5 mL PBS, pH 7.4, at 4°, add 0.3 mL of 10 mM CII in PBS, pH 7.4, dropwise. After 60 min. dilute the reaction mixture 20-fold with water, apply to a cation exchange column to separate the desired product from unreacted starting material and other products, using a linear gradient formed from equal volumes of water and 2.0 M HCl. The appropriate fractions are pooled and lyophilized to obtain the product. Alternatively, to a stirred solution of 10 mg of F or F in 5 mL PBS, pH 7.4, at 4° add 0.3 mL of 10 mM CII in PBS, pH 7.4, dropwise. After 60 min, dialyze against 3 changes of 0.5 L PBS, pH 7.4, at 4°, each for 2 hr. 4- + C 5 H 4 N-S-S-CH 2 CHa-CONH-R; where R is D, E or F. -a 2 N-tyr-lys-Pepll-CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-Pepll-CONH 2 HN-CO-CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-R where R is D (CHId) , E (CHIe) or F (Clllf ) . After derivatization of the e-N-succinyl-lys with H 2 N-CH 2 -CH 2 -S-S- CR j CHa-NH-t-BOC and deblocking, the ligand Pep2 is synthesized on the resin using standard solid phase technique. Deblocking and cleavage from the resin yields : H 2 N-tyr-lys-Pepll-CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-Pepll-CONH 2 (CIVh) CHa) 4 -NHCO- ( CR, ) 2 -CONH- ( CR, ) 2 -S-S- ( CR, ) 2 -CONH-Pep2-COOH The resin bound Lys-eNH-COf CH^ g NB- j intermediate is coupled with succinimidyl 3 (2-pyridyldithio) propionate and then deprotected and cleaved to yield: H 2 N-tyr-lys-Pepll-CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-Pepll-CONH 2 ( CR; ) 4 NHCO- (CR; ) g-NHCO-CRjCRj-S-S-CgRjN ( CV) I + Fab ' -SH HaN-tyr-lys-Pepll-CONH-CI^C^-S-S-C^C^-CONH-Pepll-CON^ (CR ) 4 NHC0- (C-_ g -NHCO-CR j CR,-S-S-Fab ' (CVIg) The addition of the nuclear localization sequence yields : H 2 N-tyr-lys-Pepll-CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-Pepll-CONH 2 ( CH 2 ) 4 NHCO- ( CH 2 ) s -NHCO-R-NH 2 where R is Pep7 ( CVIIi ) , Pep8 ( CVIIj ) , Pep9 ( CVIIk ) or PeplO ( CVII1 ) . H 2 N-tyr-lvs-Pepll-CONH-CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-Pepll-CONH 2 ( CH 2 ) 4 NHCO- ( CH 2 ) 2 -CONH-R-CONH 2 where R is Pep3 (CVIIIi), Pep4 (CVIIIj), Pep5 (CVIIIk) or Pep6 (CVIIII). Example 9 Octacationic DNA Binding Templates with Dual Ligands With at least 8 DNA binding templates and at least 12 receptor ligands, there are many possible combinations which can be used. Representative examples include polyamine templates with either a cleavable or non-cleavable spacer joining the templates and oligopeptide templates with either a cleavable or non-cleavable spacer joining the template. A schematic flow chart for the synthesis of these compounds is shown in Figure 12. The chemical pathway of synthesis is shown below. For polyamine templates: Examples 7-10 are all solid phase peptide synthesis. Final coupling are the same for each peptide as described for the A, A' , B, B' , G, G' ligands. Reaction conditions for D, E, F and the Fab' are comparable. LΣΣΣI + OH" i t-BOC t-BOC t-BOC-HN-(CH 2 ) 3 -N-CH 2 -CH-(CH 2 ) 2 -N-(CH 2 ) 3 -NH 2 (CIΣ) OCR j -COOH 4- + FMOC-C1 In reactions similar to those in Example 7 the following products are found. One skilled in the art will recognize that the starting material and resulting products are different but the reaction is the same. CΣ + H^-CHaCH^S-S-CR j CR j -NH-R where R is A, B or G. 4- t-BOC t-BOC t-BOC-HN- ( CR j ) 3 -N I -CH 2 -CH- ( CR, ) 2 -N I- ( CH 2 ) 3 -NH-FMOC OCH^COHN-CH^R j -S-S-CR j CR j -NH-R where R is A (CXIa ) , B ( CXIb ) or G ( CΣIc ) . 4. + OH~ t-BOC t-BOC t-BOC-HN- ( CR j ) 3 -N I -CR j -CH- ( CR j ) 2 -N I- ( CR j ) 3 -NH 2 OCHa-COHN-CRjCRrS-S-CRjCRj-NH-R where R is A (CΣIIa ) , B ( CΣIIb ) or G (CΣIIc ) . Dissolve 2 mmol CΣ in 2 mL dry DMF, add 4.0 mmol l-ethyl-3-[3 (dimethylamino)propyl) carbondiimide and stir 2 hr, then add 2.1 mmol N-hydroxysuccinimide and continue stirring for another 6 hr at room temperature, then combine with 2 mmol S(2-aminoethyl) S' (2-pyridyl)- dithiol in 2 mL dry DMF. After stirring overnight at room temperature, remove the solvent in vacuo, separate the product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate, 0 to 50% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product as an amorphous solid. 4- t-BOC t-BOC t-BOC-HN-(CHa) 3 -N I-CR,-CH-(CR,) 2 "N I-(CR,) 3 -NH-FMOC OCR,-COHN-CR j CR j -S-S-CgH 4 N (CΣIII) Dissolve 2 mmol CVIII in 10 mL ethanol containing 20 mmol piperidine. After overnight at room temperature, the solution is transferred to a separatory funnel, to which 50 mmol of HCl, 5 mL of water and 25 mL of benzene is added. After extraction with 3 additional portions of benzene, the combined organic phase is taken to dryness in vacuo . Separate the desired product by solid phase extraction on phenyl-silica and elution with a linear gradient of ethyl acetate to 100% in hexane. Pool the appropriate fractions and evaporate the solvent to obtain the product as an amorphous solid. t-BOC t-BOC t-BOC-HN-(CR j ) 3 -N I-CH 2 -CH-(CR,) 2 -N I-(CR,) 3 -NH 2 OCH 2 -COHN-CH 2 CH 2 -S-S-C 3 H 4 N (CΣIV) Using the same procedures for CΣIII, except substituting S-t-BOC- mercaptoethylamine for H 2 N-CH 2 CH 2 -SS-C 5 H 4 N. CΣ + H 2 N-CH 2 CH 2 -S-t-BOC 4. t-BOC t-BOC t-BOC-HN-(CR,) 3 -N I-CH 2 -CH-(CR,) 2 -N I-(CR,) 3 -NH-FMOC OCH 2 -COHN-CH 2 CH 2 -S-t-BOC (CΣV) 4- + OH ~ t-BOC t-BOC t-BOC-HN-(CH 2 ) 3 -N I-CH 2 -CH-(CR,) 2 -N I-(CH 2 ) 3 -NH 2 OCH 2 -COHN-CH 2 CH 2 -S-t-BOC (CXVI) The nuclear localization sequences are added. Standard continuous-flow solid phase synthetic methodologies are used to couple the commercially available 5-(N-t-BOC) aminohexanoic acid to the protected peptide on the solid support and the subsequent reaction to give CXVIIa-c, CΣVIII and CΣIΣ, as the final products after deprotection and release from the support and chromatographic isolation. H 2 N-Pep3-CONH-support + t-BOC-NH-(CR ; ) 5 -COOH 4. NH 2 -(CR,) 5 -CONH-Pep3-CONH-support 4- + CX 0-CR,-OC-NH- (CR,) 5 -COHN-Pep3-CONH-support t-BOC-HN- ( CHa ) 3 -N-CR j -CH- ( CH 2 ) 2 -N- ( CR j ) 3 -NH-FMOC t-BOC t-BOC 4- + OH~ 0-CH 2 -OC-NH- (CHa) 5 -COHN-Pep3-CONH-support t-BOC-HN- ( CHa) 3 -N-CR,-CH- (CR,) 2 "N- (CHa) 3 -NH 2 t-BOC t-BOC 4- + HOOC-CH 2 CH 2 -S-S-CH 2 CH 2 -COOCH 2 C 6 H 3 ( -OCH 3 ) 2 4- + H + 0-CH 2 -OC-NH-(CH 2 ) g -COHN-Pep3-CONH-support HN- ( CHa) 3 -N-CR j -CH- ( CR j ) 2 -N- (CR, ) a-NHCO-C^CHa-S-S-C^C^-COOH t-BOC t-BOC t-BOC 4. + R; Where R is CX a, CΣIIb, CΣIIc, CΣIV or CΣVI Where R is S-CR j CR j -NH-A, S-CH j CHg-NHCO-B, S-CH2CH 2 -NHCO-L, S-C 5 H 4 N or S-t- BOC 0-CH 2 -OC-NH- ( CH 2 ) 5 -COHN-Pep3 -CONH 2 H j -N- ( CR j ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CH 2 ) 3 -NHCO-CR,CR,-S H 2 N- ( CH 2 ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 -NH- ( CH 2 ) 3 -NHCO-CH 2 CH 2 -S OCH 2 -COHN-CH2CH 2 -S-R Where R is S-CH 2 CH 2 -NH-A (CΣVIIa), S-CH 2 CH 2 -NHCO-B (CXVIIb), S-CR-CR,- NHCO-G (CXVIIC), S-C 5 H 4 N (CXVIII) or SH. (CXIX) . To a stirred solution of 10 mg of Fab' -SH in 5 mL PBS, pH 7.4, at 4°, add 0.3 mL of 10 CXVIII in PBS, pH 7.4, dropwise. After 60 min, dialyze against 3 changes of 0.5 L PBS pH 7.4, at 4°, each for 2 hr. Alternatively to a stirred solution of HS-CH 2 CH 2 -CONH-Pep2-COOH, prepared by standard solid phase peptide methodology, 10 mg in 5 mL PBS, pH 7.4, at 4°, add 0.3 mL of 10 mM CXVIII in PBS, pH 7.4, dropwise. After 60 min, CXVIII + HR Where R is S-Fab' (CX∑g) or S-CR,CR,-CONH-Pep2-COOH (C∑∑h) . 4. 0-CH 2 -OC-NH- (CH 2 ) 5 -COHN-Pep3-CONH 2 R j N- ( CR, ) 3 -NH-CH 2 -CH- ( CR, ) 2 -NH- ( CR j ) g-NHCO-CH- j CR j -S H 2 N- ( CH 2 ) 3 -NH-CH 2 -CH- ( CH 2 ) 2 ~NH- ( CH 2 ) 3 -NHCO-CH 2 CH 2 -S OCH 2 -COHN-CH 2 CH 2 -S-R Where R is S-Fab' (CXΣg) or S-CR.CH 2 -CONH-Pep2-COOH (CXXh) . Using similar reaction conditions as for preparing Cllld-f CXIΣ + CgH^-S-S-CR-CI^-CONH-R where R is D, E or F. 4. 0-CR--OC-NH- ( CR j ) g -COHN-Pep3-CONR j H2N- ( CH 2 ) 3 -NH-CH 2 -CH- ( CR j ) 2 -NH- ( CH 2 ) 3 -NHCO-CH 2 CH 2 -S H 2 N- ( CR, ) 3 -NH-CH 2 -CH- ( CR ) 2 ~NH- ( CR, ) 3 -NHCO-CR j CR,-S OCHa-COHN-CR j CHa-S-R where R is D (CΣΣId) , E (CΣΣIe) or F (CXXIf ) . Substitution of succinic anhydride for HOOC-C j CH^S-S-CHaCHa-COOHa- C 6 H 3 (-OCH 3 ) 2 at the fourth stage of synthesis gives a noncleavable intermediate which is further modified according to the reaction sequences for CΣVIIa, CΣVIIb, C∑∑g, C∑∑h. CΣΣId, CΣΣ e and CΣΣIf to give the following products for gene delivery. Standard continuous-flow solid phase synthetic methodologies are used to couple succinic anhydride to the protected peptide on the solid support and the subsequent reaction to give CΣΣIIa-c, and CΣΣVh, as the final products after deprotection and release from the support and chromatographic isolation. The intermediates corresponding to CΣVIII and CΣIΣ, are deprotected, released from the support, chromatographically purified, and reacted with the appropriate intermediates to give CΣΣVg, CXXVId-f, as described for the CΣX and CΣΣ series. 0-CH 2 -OC-NH-(CR j ) 5 -COHN-Pep3-CONH 2 R j N-(CR,) g -NH-CR j -CH-(CHa) 2 "NH-(CR,) 3 -NHCO-CR j RjN-(CRj) 3 -NH-CH 2 -CH-(CHa) 2 -NH_ ( CH 2 ) 3 -NHCO-CRj OCH 2 -COHN-CH 2 CH 2 -S-R where R is S-CR-CR j -NH-A (CΣΣIIa), S-CR,CH 2 -NHCO-B (CΣΣIIb), S-CR,CH 2 - NHCO-G (CΣΣIIc), S-Fab' (CΣΣVg) , S-CHaCH 2 -CONH-Pepl-CONH2 (CΣΣVh) , S- CHaCH 2 CO-D (CΣΣVId), S-CHaCR j -CO-E (CΣΣVIe) or S-CR ϊH j -CONH-F (CΣΣVIf) Example 10 Further Octacationic DNA Binding Templates with Dual Ligands A schematic flow chart for the synthesis of these compounds is shown in Figure 13. The chemical pathway of synthesis is shown below. Examples 7-10 are all solid phase peptide synthesis. Final coupling are the same for each peptide as described for the A, A' , B, B' , G, G' ligands. Reaction conditions for D, E, F and the Fab' are comparable. For oligopeptide template t-BOC-HN-tyr-lys-Pepll-CO-support R j NOC-Pep3-HNOC-(CH 2 ) 2 -OCHN-(CH 2 ) 4 4. H + H^-tyr-lys-Pepll-CO-support R j NOC-Pep3-HNOC-(CR,) 2 -OCHN-(CR,) 4 4- + FMOC-NH-CH^H^S-S-CR j CHa-COOH FMOC-NH-CH 2 CH 2 -S-S-CH 2 CH 2 CONH-tyr-lys-Pepll-CO-support H 2 NOC-Pep3-HNOC- ( CH 2 ) 2 -OCHN- ( CH 2 ) 4 4- OH~ NH 2 -CH 2 CH 2 -S-S-CH 2 CH 2 -CONH-tyr-lys-Pepll-CO-support H 2 NOC-Pep3 -HNOC- ( CH 2 ) 2 -OCHN- ( CH 2 ) 4 Further extension of template yields : ( CH 2 ) 4 -eNH 2 S-CH 2 CH 2 -NH-CO-Pepll-lys-tyr-NH-t-BOC S-CH 2 CH 2 -CONH-tyr-lys-Pepll-CO-support TCH 2 ) 4 NHCO-(CH 2 ) 2 -CONH-Pep3-CONH 2 After derivatization of the e-N-lys with succinic anhydride the ligands (X) are coupled to the resin bound protective peptide. X where X is R j N-CR j CR j -S-S-C 8 ^N, H 2 N-CH 2 CH 2 -S-t-BOC or H 2 N-CR,CH 2 -S-S-CR,CH 2 -NH-R and where R is A, B or G. Although not necessary, it is sometimes desirable to derivatize the amino-terminal tyr with a 2-methoxy-6-chloracridinyl moiety. If not, then deprotection and release from the resin yields: Where R is S-CH 2 CH 2 -NHCO-A ( CΣΣVIIa ) , S-CR,CH 2 -NHCO-B ( CΣΣVIIb ) , S-CH 2 CH 2 - NHCO-G ( CΣΣVIIc ) , S-C 5 H 4 N ( CΣΣIΣ ) or SH ( CΣΣΣ ) . CXXΣ + C g R j N-S-S-CH jj CH jj -CONH-R; where R is D , E or F 4. Where R is D (CΣΣΣId), E (CΣΣΣIe) or F (CΣΣΣIf) After derivatization of the e-N-succinyl-lys with H g N-C^C^-S-S- CH g CHa-NH-t-BOC and deblocking the ligand Pep3 is synthesized on the resin using standard solid phase techniques. Deblocking and cleavage from the resin yields: HOOC-Pep2-HNOC-CH 2 CR j -S (CHa) 4 eNH-CO-CR ; CR j -CONH-CH 2 CR j -S S-CR j -CR j -NH-CO-Pepll-lys-tyr-NR, I (CΣΣΣIIh) S-CR j -CR j -CONH-tyr-lys-Pepll-COOH ( CR, ) 4 NHCO- ( CHa ) 2 -CONH-Pep3 -CONH 2 CΣΣΣIΣ + Fab' -SH 4- Fab' -R-HNOC-CH 2 CHa-S ( CR, ) 4 eNH-CO-CH 2 CH 2 -CONH-CH 2 CH 2 -S S-CR j -CR j -NH-CO-Pepll-lys-tyr-NR, I ( CΣΣΣIIIg) S-CHa-CHa-CONH-tyr-lys-Pepll-COOH (CR j ) 4 NHC0-(CR j ) 2 -CONH-Pep3-CO Ha Substitution oft-BOC-NH-(CR j ) 5 -COOH for FMOC-NH-CHaCHa-S-S-C j -CHa- COOH at the second stage of synthesis gives a noncleavable intermediate, which is further modified according to the reaction sequence for the CΣΣVII series, C∑∑∑g, CΣΣΣIIIh and the CΣXX series to give the following products for gene delivery. R-HNOC-CR j CHa-S ( CHa ) 4 eNH-CO-CR,CR,-CONH-CR j CH 2 -S CHa- ( CH 2 ) 4 -NH-CO-Pepl 1- ly s-tyr-NR, CONH-tyr-l Lyy rss -Pepll-COOH ( CH 2 ) 4 NHCO- ( CH 2 ) 2 -CONH-Pep3 -CON-^ where R is S-CH 2 CH 2 -NH-A (CΣΣΣIVa), S-CH 2 CH 2 -NHCO-B (CΣΣΣIVb), S-CH 2 CH 2 - NHCO- (CΣΣΣIVc), S-Fab' (CΣΣΣIΣg), S-CR,CH 2 -CONH-Pepl-CONH 2 (CΣΣΣVIIIh) , S-CH 2 CH 2 .CO-D (CΣΣΣVIId), S-CH 2 CH 2 -CO-E (CΣΣΣVIIe) , S-CR,CH 2 -CONH-F (CΣΣXVIIf) . Example 11 Oligonucleotides Containing Receptor Ligands Figure 14 show a double stranded DNA vector with a single stranded DNA attached to a ligand containing both a plasma membrane and nuclear membrane receptors. Two different functions can be shown for this single stranded pyrimidine deoxyoligonucleotide modified at the 3' and 5' terminal nucleotides with a space molecule derivatized with a ligand for either a plasma membrane receptor or nuclear membrane receptor and/or a nucleotide containing a modified base conjugated with a ligand for either a plasma membrane receptor or a nuclear membrane receptor. The first function is to target double stranded vectors to specific cells and then to the nucleus of the targeted cell for expression of the vector and/or integration of the vector sequences into the host genome. One to ten copies of the double stranded target sequences either individually or clustered will be inserted in non- coding regions of the vector. The second function is to deliver therapeutic single stranded DNA for treatment of cancer, infectious disease and cardiovascular disease. Targeting the single stranded DNA to specific cells and then to the nucleus of the targeted cell will form a triplex structure that prevents transcription of the specified genes. In Figure 15A is shown single stranded DNA as a DNA-binding template containing a single receptor ligand. C m is the 5-methyl cytosine derivative. In Figure 15B is shown single stranded DNA-binding template in which N-(2-ethylamino)glycine replaces the deoxyribose-phosphate backbone of the nucleic acid polymer. Derivatives and analogs of Figure 15 are shown in Figures 15C. In Figure 15D is an example of a ligand containing template in the pyrimidine series. With at least 18 binding templates and at least 12 receptor ligands there are many possible combinations for use. It is obvious to one skilled in the art that two different DNA binding templates could be linked 5' (3' ) to 3' (5' ) with a dithio bridge so that the single stranded DNA bearing the plasma membrane ligand would disassociate from the double stranded DNA vector. The oligonucleotides are made by conventional solid phase synthesis. The 5' and 3' nucleotides contained in an amino group in lieu of the 5' and 3' hydroxyl moieties, respectively, of the terminal nucleotides. The nucleotide T-ϊ is 5-(N-[N-{N-ligand-5-aminohexanoyl}-4-aminobutanoyl]- 3-aminoallyl]-2'-deoxyuridine moiety. The nucleotide A-Y is 8-[N-[N- ligand-5-aminohexanoyl]-8-aminohexylamino]-2'-deoxyadenosine moiety. The ligand for SV-40 sequences is shown in Figure 16. Therapeutic single stranded DNA for the treatment of cancer infectious disease and cardiovascular disease can be delivered to specific cells and then to the nucleus of the targeted cell where it forms triplex structures that prevent transcription of the specified genes. An initial template will contain two different single stranded DNA templates linked 5' (3' ) to 3' (5' ) with a dithio bridge, so that as a result of reduction in the cytoplasm, both the single stranded DNA and the plasma membrane ligand will disassociate from the double stranded DNA vectors as separate molecules. The spacer for the plasma membrane ligand also contains a dithio moiety so that the cellular targeting ligand will be released when the complex is present in the cytoplasm. The oligonucleotides are made by conventional solid phase synthesis. The 5' and 3' nucleotides contain an amino group in lieu of the 5' and 3' hydroxyl moiety, respectively, of the terminal nucleotides. Figure 17 shows an example of this for the C-myc promoter. In Figure 17 the deoxyoligonucleotide strands are synthesized on an automated DNA synthesizer using a solid-phase cyanoethylphosphoramidate method. Commercially available reagents are used to provide a 3' terminal thiol which is reacted with further reacted A (see Example 1) after deblocking and release of the oligonucieotide from the support. Dissolve 2 mmol of the protected peptide N-succinyl-Pep5-CONR, released from the peptide support in 2 mL dry DMF, and add 4.0 mmol l-ethyl-3- [3- (dimethylamino)propyl)carbodiimide. Stir for 2 hr, then add 2.1 mmol N-hydroxysuccinimide. Stir for another 6 hr at room temperature, then couple with the deprotected side chain amino group of 5-(N-[N-{N- ligand-5-aminohexanoyl}-4-aminobutanoyl]-3-aminoallyl)-2'-deoxyuridine moiety on the solid support. The nucleotide is terminated with the commercially available Uni-Link AminoModifier (Clontech) . The terminal Fmoc amino protecting group be removed and reacted with 6,9-dichloro-2- methoxyacridine before the substituted oligonucieotide is cleaved from the support. All patents and publications mentioned in the specification are indicative of the levels of those skilled in the art to which the invention pertains. All patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference. One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned as well as those inherent therein. The DNA transporter systems along with the methods, procedures, treatments, molecules, specific compounds described herein are presently representative of preferred embodiments are exemplary and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art which are encompassed within the spirit of the invention are defined by the scope of the claims. What we claim is:

Claims: Claims 1. A DNA transporter system for inserting specific DNA into a cell, comprising: a plurality of a first DNA binding complex, said complex including a first binding molecule capable of non-covalently binding to DNA, said first binding molecule covalently linked to a surface ligand, said ligand capable of binding to a cell surface receptor; a plurality of a second DNA binding complex, said complex including a second binding molecule capable of non-covalently binding to DNA, said second binding molecule covalently linked to a nuclear ligand, said nuclear ligand capable of recognizing and transporting the transporter system through a nuclear membrane; wherein said plurality of first and second DNA binding complexes are capable of simultaneously, non-covalently binding to the specific DNA. 2. The transporter of claim 1 wherein, the first binding molecule is selected from the group consisting of spermine, spermine derivative, histones, cationic peptides and polylysine; and the second binding molecule is selected from the group consisting of spermine, spermine derivative, histones, cationic peptides and polylysine. 3. The transporter of claim 2, wherein the first and second binding molecules are the same molecule and are a spermine derivative. 4. The transporter of claim 1, wherein the surface ligand is a molecule which binds to a receptor selected from the group consisting of folate receptor, biotin receptor, lipoic acid receptor, low density lipoprotein receptor, asialoglycoprotein receptor, insulin-like growth factor type II/cation-independent mannose-6-phosphate receptor, calcitonin gene-related peptide receptor, insulin-like growth factor I receptor, nicotinic acetylcholine receptor, hepatocyte growth factor receptor, endothelin receptors or bile acid receptor. 5. The transporter of claim 1, wherein the surface ligand is a molecule which binds to an IgG antigen. 6. The transporter of claim 1, wherein the surface ligand includes a compound selected from the group consisting of Pepl, Pep2, Pepl2, Pepl3, Pepl4, Pepl5, Pepl6, Pepl7, Pepl8, Pepl9, Pep20, Pep21, Pep22, Pep23, A, B, G, D, E, P, J, M and Fab' 7. The transporter of claim 1, wherein the nuclear ligand is a molecule selected from the group consisting of Pep3, Pep4, Pep5, Pep6, Pep7, Pep8, Pep9 and PeplO. 8. The transporter of claim 1, wherein the surface ligand and nuclear ligand are attached to their respective binding molecule by a spacer. 9. The transporter of claim 8, wherein the spacer is hydrophilic and has from 6-30 carbons. 10. The transporter of claim 9, wherein the spacer has from 6-16 carbons. 11. The transporter of claim 9, wherein the spacer is a repeating ω-amino acid of the structure [NH-(CH 2 -CH 2 ) n -CO-] m wherein n = 1-3 and m = 1-20, 12. The transporter of claim 9, wherein the spacer is a disulfide of the structure (CR,CR,-S-S-CH 2 CH 2 -) n 13. The transporter of claim 19, wherein the spacer is an acid sensitive bifunctional molecule of the structure -CO-CH 2 -C = CH-NH-CR,-CH 2 -S- COOH 14. The transporter of claim 1, further comprising a plurality of a third DNA binding complex, said complex including a third binding molecule capable of non-covalently binding to DNA, said third binding molecule covalently linked to a virus or a lytic peptide; wherein said plurality of third DNA binding complexes are capable of simultaneously, non-covalently binding to said specific DNA. 15. The transporter of claim 14 wherein, the third binding molecule is selected from the group consisting of spermine, spermine derivative, histones, cationic polypeptide and polylysine. 16. The transporter of claim 14, wherein the third binding molecule is spermine derivative. 17. The transporter of claim 14, wherein the surface ligand is a molecule which binds to a receptor selected from the group consisting of folate receptor, biotin receptor, lipoic acid receptor, low density lipoprotein receptor and asialoglycoprotein receptor, insulin-like growth factor type II/cation-independent mannose-6-phosρhate receptor, calcitonin gene-related peptide receptor, insulin-like growth factor I receptor, nicotinic acetylcholine receptor, hepatocyte growth factor receptor, endothelin receptors or bile acid receptor. 18. The transporter of claim 1, wherein the surface ligand is a molecule which binds to an IgG antigen. 19. The transporter of claim 1, wherein the surface ligand includes a compound selected from the group consisting of Pepl, Pep2, Pepl2, Pepl3, Pepl4, Pepl5, Pepl6, Pepl7, Peplδ, Pepl9, Pep20, Pep21, Pep22, Pep23, A, B, G, D, E, P, J, M and Fab' . 20. The transporter of claim 14, wherein the nuclear ligand is a molecule selected from the group consisting of Pep3, Pep4, Pep5, Pep6, Pep7, Pep8, Pep9 and PeplO. 21. The transporter of claim 14, wherein the virus is selected from the group consisting of adenovirus, parainfluenza virus, herpes virus, retrovirus and hepatitis virus. 22. The transporter of claim 14, wherein the lytic peptide is selected from the group consisting of Pep24, Pep25 and Pep26. 23. The transporter of claim 14, wherein the virus is adenovirus. 24. The transporter of claim 14, wherein the virus ligand is attached to the binding molecule by a spacer. 25. The transporter of claim 24, wherein the spacer is hydrophilic and has from 6-30 carbons. 26. The transporter of claim 25, wherein the spacer has from 6-16 carbons. 27. The transporter of claim 25, wherein the spacer is a repeating ω-amino acids having a structure of [NH-(CR,-CR,) n -C0-] m where n = 1-3 and m = 1-20, 28. The transporter of claim 25, wherein the spacer is a disulfide having the structure of (CI^CR J -S-S-CR J -CRJ-) . 29. The transporter of claim 14, wherein the spacer is an acid sensitive bifunctional molelcule of the structure -CO-CR,-C = CH-NH-CR,-CR,-S- COOH 30. The transporter of claim 14, wherein the lytic peptide is adenovirus. 31. The transporter of claim 14, wherein the lytic peptide ligand is attached to the binding molecule by a spacer. 32. The transporter of claim 24, wherein the spacer is hydrophilic and has from 6-30 carbons. 33. The transporter of claim 25, wherein the spacer has from 6-16 carbons. 34. The transporter of claim 25, wherein the spacer is a repeating ω-amino acids having a structure of [NH-(CH 2 -CH 2 ) n -CO-] m where n = 1-3 and m = 1-20, 35. The transporter of claim 25, wherein the spacer is a disulfide having the structure of (CR ; CH 2 -S-S-CH 2 -CH 2 -) . 36. The transporter of claim 14, wherein the spacer is an acid sensitive bifunctional molelcule of the structure -C0-CH 2 -C = CH-NH-CH 2 -CH 2 -S- COOH 37. A DNA transporter for inserting specific DNA into a cell, comprising a plurality of a first DNA binding complex, said complex including a first binding molecule capable of non-covalently binding to DNA, said first binding molecule attached to a first spacer, said first spacer also attached to a surface ligand, said ligand capable of binding to a cell surface receptor; a plurality of a second DNA binding complex, said complex including a second binding molecule capable of non-covalently binding to DNA, said second binding molecule attached to a second spacer, said second spacer also attached to a nuclear ligand capable of recognizing and transporting the transporter system through a nuclear membrane; a plurality of a third DNA binding complex, said complex including a third .binding molecule capable of non-covalently binding to DNA, said third binding molecule attached to a third spacer, said third spacer also attached to a virus or lytic peptide; wherein said plurality of first, second and third DNA binding complexes are capable of simultaneously, non-covalently binding to the specific DNA. 38. The DNA transporter of claim 37, wherein the first, second and third binding molecules are a spermine derivative; the surface ligand is a molecule which binds to the asialoglycoprotein; the nuclear ligand is selected from the group consisting of Pap3, Pep4, Pep5, Pep6, Pep7, Pep8, Pep9 and PeplO; the virus is the adenovirus; the first, second and third spacer is hydrophilic and contains 6-16 carbons. 39. A DNA transporter system for inserting specific DNA into a cell, comprising: a plurality of a common DNA binding complex, each of said complexes includes a binding molecule capable of non-covalently binding to DNA, said binding molecule attached to both a surface ligand capable of binding to a cell surface receptor and a nuclear receptor capable of recognizing and transporting the transporter system though a nuclear membrane. 40. The transporter of claim 39, further comprising a spacer linking the surface ligand and nuclear ligand to the binding molecule. 41. The transporter of claim 39, further comprising a virus attached to said binding molecule. 42. The transporter of claim 41, further comprising a spacer linking , the virus to the binding molecule. 43. A method of introducing DNA into a cell comprising the step of contacting the cell with the DNA transporter of claim 1. 44. A method for in vivo targeting of the insertion of DNA into a cell comprising the step of contacting the cell with the DNA transporter of claim 1, wherein the surface receptor is specific to the cell which is targeted. 45. A method for prevention or treatment of disease comprising the step of contacting a cell within an organism with a therapeutic dose of the DNA transporter of claim 1, wherein the specific DNA includes a molecule to treat the disease. 46. The method of claim 45, wherein the organism is a human. 47. The method of claim 45, wherein the organism is an animal. 48. A method for modifying animals comprising the step of contacting a cell within the animal with a DNA transporter of claim 1, wherein the specific DNA includes the sequence to modify the animal. 49. A compound selected from the group consisting of Xa, Xb, Xc, XIa, ∑lb, X c, XΣΣIΣa, ΣΣΣIXb, XΣΣIΣc, Ll∑a, Ll∑b, LIΣc, LΣXXVIIa, LΣΣΣVIIb and LXXXVIIc. 50. A compound selected from the group consisting of Xllld, ∑llle, XLIId, XLIIe, LXIId, LΣIIe, ΣC d and ΣCIe. 51. A compound selected from the group consisting of ∑lllf, ΣLIIf, LΣIIf and ΣCIf. 52. A compound selected from the group consisting of XVIlg, XLVIg, LΣVIg and ΣCVg. 53. A compound selected from the group consisting of ΣΣ Σg, LΣΣVIIIg and CVIg. 54. A compound selected from the group consisting of ΣΣVIIh, LΣΣVIh and CIVh. 55. A compound selected from the group consisting of ΣΣI i, ΣΣIIj, ΣΣIIk, ΣΣII1, Lli, LIj, LIk, LII, LΣΣIi, L ΣIj, LΣΣIk, LΣΣI1, ΣCΣi, ΣCΣj, ΣCΣk and ΣCΣ1. 56. A compound selected from the group consisting of ∑l∑h, ΣLVIIIh, LΣVIIh and ΣCVIIh. 57. A compound selected from the group consisting of ∑∑∑i, ΣΣΣj, ∑∑∑k, ∑∑∑l, XXΣIi, ΣΣΣIj, XXΣIk, ΣΣΣI1, LΣΣIΣi, LΣΣIΣj, LΣΣIΣk, LΣΣIΣ1, LXXXi, L∑∑∑j, LΣXXk, LXΣΣ1, CVIIi, CVIIj, CVIIk, CVIIl, CVIIIi, CVIIIj, CVIIIk and CVIIII. 58. A compound selected from the group consisting of A, A' , B, B' , G, G' , and M. 59. A compound of the structure of F. 60. A compound of the structure E. 61. A compound of the structure D. 62. A compound selected from the group consisting of CΣLIi. XLIj, CΣLIk, CΣLI1, CΣLIIi, CΣL Ij, CΣLIIk and CΣLII1. 63. A compound of the structure CΣLIII. 64. A compound selected from the group consisting of XVIII, XLVII, LXVII, ΣCVI and CΣIII. 65. A compound of the structure CΣIΣ. 66. A compound selected from the group consisting of CΣLak, CΣLbk and CΣLck. 67. A compound selected from the group consisting of CΣLdk and CΣLek. 68. A compound of the structure CΣLgk. 69. A compound of the structure CXLhk. 70. A compound of the structure CXLfk. 71. A compound selected from the group consisting of CXVIIa, CΣVIIb, CΣVIIc, CΣΣIIa, CΣΣIIb and CXΣIIc. 72. A compound of the structure C∑∑g or CΣXVg. 73. A compound of the structure CXXh or CΣXVh. 74. A compound selected from the group consisting of CΣXId, CXΣIe, CΣΣVId and CXΣVIe. 75. A compound of the structure CΣΣIf or CXXVIf. 76. A compound selected from the group consisting of ΣΣVId, ΣΣVIe, LΣΣVd, LΣΣVe, CIIId and Cllle. 77. A compound selected from the group consisting of ΣΣVIf, LΣΣVf and Clllf. 78. A compound selected from the group consisting of ΣVI, LΣV, ΣCIV and CΣVI. 79. A compound selected from the group consisting of IV, XXXIII, XI, XII, XL, XLI, LX, LΣI, LΣΣΣVIII and CΣ. 80. A compound selected from the group consisting of XV, LXIV, XCIII, ΣLV and CΣIV. 81. A compound selected from the group consisting of ΣΣIIIa, XXIIlb, ΣΣIIIc, LΣΣIIa, LΣΣIIb, LΣΣIIc, Cla, Clb and CIc. 82. A compound selected from the group consisting of ΣΣIV, ΣΣV, LΣΣIII, LΣΣIV and CII. 83. A compound selected from the group consisting of VII and XXXVI. 84. A compound selected from the group consisting of ΣΣVIII, LΣΣVII and CV. 85. A compound selected from the group consisting of LIV, LVI, LXXΣII, LΣΣΣIV and CΣ. 86. A compound selected from the group consisting of ΣΣI, L, LΣΣ and ΣCIΣ. 87. A compound selected from the group consisting of CΣΣVIIa, CXΣVIIb, CΣXVIIc, CXΣΣIVa, CΣΣΣIVb and CΣΣΣIVc. 88. A compound selected from the group consisting of CΣΣΣId, CΣΣΣIe, CΣΣΣVIId and CΣΣΣVIIe. 89. A compound selected from the group consisting of CΣΣΣIf and CΣΣΣVIIf. 90. A compound selected from the group consisting of CΣΣΣIXg and CXΣΣIIIg. 91. A compound selected from the group consisting of CXXXIIh and CΣΣΣVIIIh. 92. A compound of the structure CΣΣΣ. 93. A compound of the structure CΣΣIΣ. 94. The transporter of claim 1, wherein the plurality of first DNA binding complex are selected from the group consisting of Xa, Xb, Xc, ∑la, ∑lb, ΣIc, XXXIXa, XXXIXb, XΣΣIΣc, Ll∑a, Ll∑b, LIΣc, LΣΣΣVIIa, LXΣΣVIIb, LΣΣΣVIIc, ΣHId, ΣHIe, ΣLIId, ΣLIIe, LΣIId, LΣIIe, ΣCId, ΣCIe, ΣVIIg, ΣLVIg, LΣVIg, ΣCVg, Σ IΣg, LΣΣVIIIg, CVIg, ΣΣVIIh, LΣΣVIh, CIVh, XlXh, XLVIIIh, LΣVIIh, ΣCVIIh, CΣLak, CXLbk, CΣLck, CXLdk, CΣLek, CΣLgk, CΣLhk, CXLfk, ΣΣVId, ΣΣVIe, LΣΣVd, LΣΣVe, CHId, Cllle, ΣΣIIIa, ΣΣIIIb, ΣΣIIIc, LΣΣIIa, LΣΣIIb, LΣΣIIc, Cla, Clb, CIc and combinations thereof. 95. The transporter of claim 1, wherein the plurality of second DNA binding complex are selected from the group consisting of ΣΣIIi, ΣΣIIj, ΣΣIIk, ΣΣII1, Lli, LIj, LIk, LII, LΣΣIi, LΣΣIj, LΣΣIk, LΣΣI1, ΣCΣi, ΣCΣj, ΣCΣk, ΣCΣ1, ∑∑∑i, ∑∑∑j, ∑∑∑k, ∑∑∑l, ΣΣΣIi, ΣΣΣIj, ΣΣΣIk, ΣΣΣI1, LΣΣIΣi, LΣΣIΣj, LΣΣIΣk, LΣΣIΣl, L∑∑∑i, LΣΣΣj, L∑∑∑k, L∑∑∑l, CVIIi, CVIIj, CVIIk, CVIIl, CVIIIi, CVIIIj, CVIIIk, CVIIII, CΣLIi, CΣLIj, CΣLIk, CΣLI1, CΣLIIi, CΣLIIj, CΣLIIk, CΣLII1 and combinations thereof. 96. The transporter of claim 14, wherein the plurality of third DNA binding complex are selected from the group consisting of ΣHI , XLIIf, LXIIf, ΣCIf, CΣΣIf, CΣΣVIf, ΣΣVIf, LΣΣVf, Clllf and combinations thereof. 97. The transporter of claim 2, wherein the first an second binding molecules are selected from the group consisting of IV, XXXIII, XI, XII, XL, XLI, LX, LXI, LΣΣΣVIII, CΣ, ΣΣIV, ΣΣV, LΣΣIII, LΣΣIV, CII and combinations thereof. 98. The transporter of claim 39, wherein the plurality of common DNA complex is selected from the group consisting of CΣVIIa, CΣVIIb, CΣVIIc, CΣΣIIa, CXΣIIb, CXΣIIc, C∑∑g, CΣΣVg, C∑∑h, CΣΣVh, CΣΣId, CΣΣIe, CΣΣVId, CΣΣVIe, CΣΣVIIa, CΣΣVIIb, CΣΣVIIc, CΣΣΣIVa, CΣΣΣIVb, CΣΣΣIVc, CΣΣΣId, CΣΣΣIe, CΣΣΣVIId, CXΣΣVIIe, CXΣΣIf, CΣΣΣVIIf, CXΣΣIΣg, CΣΣΣIIIg, CXXXIIh, CΣΣΣVIIIh and combinations thereof. 99. A compound of the structure of P. 100. A compound of the structure of J. 101. A compound of the structure of Q. 102. A compound of the structure shown in Figure 15B. 103. A compound of the structure shown in Figure 15C. 104. A compound of the structure shown in Figure 15D. 105. A compound of the structure shown in Figure 22. 106. A compound of the structure shown in Figure 24. 107. A compound of the structure selected from the group consisting of 4, 10, 11, 12, 13, 14, 15 and 16. 108. A compound of the structure selected from the group consisting of 17, 18, 19 and 20. 109. A compound of the structure selected from the group consisting of 6, 21, 22, 23 and 24. 110. A compound of the structure selected from the group consisting of 25, 26 and 27. 111. A compound of the structure selected from the group consisting of 28, 29 and 30.

Inventors: SMITH LOUIS C, WOO SAVIO L C, SMITH, LOUIS, C., WOO, SAVIO, L., C.
Applicants: BAYLOR COLLEGE MEDICINE, BAYLOR COLLEGE OF MEDICINE
Application year: 1993
Main classification: C12N5
IPC codes: A61K38, A61K47, A61K48, C07H21, C07K7, C07K14, C12M1, C12N15