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Although this film was made in late 1941, it was not released until September 1944 because of a contractual obligation between Warner Bros. and the producers of the Broadway show, in which Warner Bros. agreed not to release the film until the end of the stage play's run. According to Warner Bros. press releases, Warner executive Hal B. Wallis outbid "every major film company" in February 1941 for the rights to Kesselring's play. Warner Bros. studio records included in the Warner Bros. collection at the USC Cinema-Television Archives reveal that the rights cost $175,000, and that theatrical producers Howard Lindsay and Russel Crouse negotiated for 15% of the film's profits. Although the original projected release date of the film was September 30, 1942, the play had 1,444 performances and ran for over three and a half years, thus delaying considerably the film's release. Josephine Hull, Jean Adair and John Alexander re-created their Broadway stage roles for this picture. Upon completion of their film duties, the actors then returned to the play.
Although not cast in the film, Boris Karloff originated the role of "Jonathan" on the stage. According to modern sources, Karloff volunteered to stay with the play to appease Crouse and Lindsay, who were concerned that the loss of all of their stars at one time would hurt their ticket sales. Studio records indicate that Warner Bros. suggested Humphrey Bogart to Lindsay and Crouse as a possible stage replacement for Karloff, but apparently the deal was never pursued. During filming, Karloff signed an agreement allowing the use of his name and likeness in the picture, a legal matter that greatly concerned Warner Bros. executives. In the spring of 1941, French film director Ren Clair saw the Broadway production of Arsenic and Old Lace and approached Lindsay and Crouse about directing the film. Lindsay and Crouse then communicated Clair's interest to Warner Bros., but it is not known if the studio ever seriously considered Clair for the job. Modern sources claim that Capra originally wanted Bob Hope for the role of "Mortimer," but Hope was not available.
According to studio records, Warner Bros. borrowed Cary Grant from Columbia for the production. Grant's total salary was $160,000. As per Grant's instructions, $50,000 of that money went to the Hollywood Division of the British War Relief Association of Southern California, $25,000 went to the American Red Cross, $25,000 went the United Service Organization, and $10,000 was paid to Grant's agent. Capra received $100,000 for his services. Hull and Adair were each paid $10,000, Lorre received $13,000 and Massey, $25,000. Studio records additionally record the following information about the production: For the film, the Epsteins expanded the role of "Mortimer" to accommodate Grant's star status and also added a few scenes, including an Ebbets Field baseball riot, to the beginning of the story. Prior to production, the script was submitted to the PCA for approval. According to files in the MPAA/PCA Collection at the AMPAS Library, the PCA advised Warner Bros. research head Dr. Herman Lissauer to play down the newlyweds' "sex frustration," which was evident, in their opinion, in certain lines and bits of business. In addition, the PCA strongly suggested that all references to actual poisons, with the exception of arsenic, be eliminated from the script, as an actual "recipe" for a toxic additive might be replicated by unstable viewers. Some of the PCA's advice was taken, while other suggestions were ignored without apparent ramifications.
Throughout most of the production, the script was being re-written by the Epstein brothers. Contrary to some modern sources, which state that the film was shot in four weeks, the proposed shooting period was eight weeks, and actual filming took nine weeks. (In his autobiography, Capra claims that he submitted a budget to Warner Bros. studio head Jack L. Warner that was based on a four-week shooting schedule, but no evidence that Warner Bros. ever seriously considered filming the project in that time has been found.) In a memorandum to an executive at Warner Bros., unit manager Eric Stacey described Capra's directing methods to explain why, in part, the production fell behind schedule: "He times his work so that the last thing in the day he will stage, and photograph, [is] a master long shot of a sequence; then the following morning at 8:00 o'clock when he sees his dailies of the previous day's work, he will continue in that sequence and make changes while he is shooting closer shots of the same action, and as ideas develop he will sometimes go back and make close-ups again, having the character read a different line, or have the character do a different piece of business." On November 1, 1941, a decision was made to shoot all of the Brewster house interiors, the bulk of the story, in sequence. Production records indicate that some scenes, such as ones in Mortimer's grandfather's laboratory, in the Brewster cellar, and in the aunts' bedroom, were shot but not used in the final film.
Modern sources note that a scene at the end of the story, in which "Mr. Witherspoon," played by Edward Everett Horton, becomes the aunts' last victim, was shot and included in preview prints of the film. Because of poor audience reaction to the screen demise of the popular character actor, however, the scene was removed from release prints. Publicity items from 1944 state that Massey's makeup, which was the subject of much debate during the production, required two hours to apply and two hours to remove. The production ended a few days after the United States' entry into World War II. According to a December 13, 1941 New York Times article on Capra, earlier in the year, he had applied for a commission with the Signal Corps. Shortly after finishing Arsenic and Old Lace, he assumed his duties as a major and made no other commercial films until 1946's It's a Wonderful Life, which he produced after his release from the service. Warner Bros. press releases boasted that the set for Arsenic and Old Lace was the largest ever constructed at the studio, and that the house was "complete in every detail, room by room." Because of strict movie censorship rules, the word "bastard," which is used in the stage play in a key line at the end, was not included in the screenplay. In his autobiography, Capra credited Jesse Hibbs (not Russ Saunders) as the assistant director.
Capra also notes that while he was stationed in London in 1943, he overheard American and British soldiers screaming "Charge!" in the manner of the "Teddy Roosevelt" character and deduced that they had seen the film. He then learned that Jack L. Warner had released the picture to the armed forces a year before it was to be released to the general public. An AMPAS notice indicates that Arsenic and Old Lace was not given Academy Award consideration for the fourteenth, fifteenth or sixteenth annual awards because of its 1944 release date. Daily Variety lists the film's preview running time as 97 minutes, but this time is most likely an error. In November 1946, Karloff played "Jonathan" in an abbreviated CBS radio version of Kesselring's play. Arsenic and Old Lace was produced on television four times: the first broadcast on CBS in 1945 starred Josephine Hull and Boris Karloff; Karloff and John Alexander reprised their roles for a 1955 CBS broadcast, which also starred Helen Hayes, Orson Bean and Billie Burke; in 1962, a third version, which starred Karloff, Tony Randall and Mildred Natwick, was broadcast on NBC; Hayes reprised her 1955 role for a fourth broadcast on ABC in 1969, which also starred Lillian Gish, Fred Gwynne and Bob Crane.